Composition for improving cognitive function, comprising phospholipid mixture, neutral lipids, and sphingolipids

Abstract

The present invention relates to a composition for improving cognitive function, the composition improving cognitive function by comprising: a mixture of phospholipids comprising phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, glycosphingolipids, and gangliosides; one or more types of neutral lipids selected from the group consisting of EPA and DHA; sphingolipids; and oil.

Description

A composition for improving cognitive function comprising a phospholipid mixture, a neutral lipid, and a spinolipid

[0001] The present invention relates to a composition capable of improving cognitive function, comprising a phospholipid mixture, a neutral lipid, and a spinolipid.

[0002] The human brain is an important part that controls learning, memory, movement, etc., and nerve cells are connected to each other to form a huge neural network, which becomes the nervous system foundation that includes learning and memory.

[0003] However, when harmful reactive oxygen species are excessively produced due to various causes and accumulate in the body, they cause oxidative damage to cells, leading to cell degeneration and cell necrosis. In normal cases, they are removed by the body's removal mechanisms such as superoxide dismutase (SOD), catalase, carotenoids, and glutathione, but ROS remaining in the body induce degeneration and death of brain nerve cells, negatively affecting the formation of proper neural networks, reducing learning ability and memory, and in severe cases, causing degenerative brain diseases such as Parkinson's disease and Alzheimer's disease.

[0004] Furthermore, cognitive abilities in the brain, such as learning and memory, are manifested by neurotransmitters such as acetylcholine (ACh). Secreted from nerve cell terminals, ACh normally transmits signals through synaptic receptors in response to external stimuli, such as learning. It is then broken down into acetate and choline by acetycholinesterase (AChE), which are then partially reabsorbed. This process is called the cholinergic system, and it is known that most Alzheimer's patients experience serious problems with this process. A decrease in ACh impairs the brain's cognitive abilities, preventing it from functioning normally.

[0005] Therefore, AChE inhibitors are widely used as a treatment for Alzheimer's patients. Furthermore, hypertension, a cerebrovascular disease that causes stroke and arteriosclerosis, is induced by angiotensin II. Angiotensin II is an enzyme that causes hypertension, formed by the conversion of angiotensin I converting enzyme (ACE) to angiotensin II, which is broken down by renin. Angiotensin II increases blood pressure by degrading bradykinin, an enzyme that constricts arterial blood vessels and dilates them. Furthermore, stress can also cause hypertension.

[0006] The purpose of the present invention is to provide a composition capable of improving cognitive function, comprising a phospholipid mixture, a neutral lipid, and a spinolipid.

[0007] In addition, another object of the present invention is to provide a food composition for improving brain function comprising the cognitive function composition.

[0008] In addition, another object of the present invention is to provide a pharmaceutical composition for preventing and improving Alzheimer's disease, which comprises the cognitive function composition as an active ingredient.

[0009] In addition, another object of the present invention is to provide a method for treating cognitive dysfunction using the cognitive function composition.

[0010] The cognitive function improvement composition of the present invention for achieving the above-mentioned purpose may include a phospholipid mixture including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, glycosphingolipids and gangliosides; a neutral lipid including at least one selected from the group consisting of EPA and DHA; a sphingolipide; and an oil.

[0011] The above composition may contain 40 to 70 parts by weight of neutral lipid, 20 to 50 parts by weight of spinolipid, and 130 to 160 parts by weight of fat per 100 parts by weight of phospholipid mixture.

[0012] The above phospholipid mixture may include 80 to 95 parts by weight of phosphatidylethanolamine, 30 to 50 parts by weight of phosphatidylserine, 20 to 40 parts by weight of phosphatidylinositol, 50 to 80 parts by weight of sphingomyelin, 80 to 95 parts by weight of glycosphingolipids, and 5 to 20 parts by weight of gangliosides, per 100 parts by weight of phosphatidylcholine.

[0013] In addition, the food composition for improving brain function of the present invention for achieving the other purposes mentioned above may include the cognitive function composition.

[0014] In addition, the pharmaceutical composition for preventing and improving Alzheimer's disease of the present invention to achieve another purpose described above may include the cognitive function composition.

[0015] In addition, the method for treating cognitive dysfunction of the present invention for achieving another purpose described above may include a step of administering the cognitive function composition to a subject in need thereof.

[0016] The above cognitive impairment may be at least one selected from the group consisting of dementia induced by chronic ultraviolet exposure, Alzheimer's disease, Huntington's disease, vascular dementia, ischemic stroke, traumatic brain injury, amnesia, Parkinson's disease, Pick disease, Creutzfeldt-Jakob disease, and mild cognitive impairment.

[0017] The related symptoms of the above cognitive dysfunction may be one selected from the group consisting of decreased learning ability, decreased memory, lethargy, decreased attention, depression, decreased hearing, analgesia, ahidrosis, and decreased discrimination.

[0018] The cognitive function improvement composition of the present invention exhibits neuronal cell proliferation and neuronal cell protection effects, and shows a high retention time and excellent movement tendency in the passive avoidance test and the spatial memory test (Y-maze test), confirming its effectiveness in improving cognitive function.

[0019] Accordingly, the cognitive function improvement composition of the present invention can be used not only as a food composition for improving brain function but also as a pharmaceutical composition for preventing and improving Alzheimer's disease.

[0020] Figure 1 is a graph showing the cell survival rate of hippocampal cells (HT-22 cells) when treated with a cognitive function improvement composition (a) manufactured according to Example 1 of the present invention, a cognitive function improvement composition of control group 1 (b), and a cognitive function improvement composition of control group 2 (c).

[0021] Figure 2 is a graph showing cell survival rates after treatment with 10 uM of beta-amyloid, the cognitive function improvement composition manufactured according to each Example 1, and the cognitive function improvement compositions of Control Group 1 and Control Group 2.

[0022] Figure 3 is a schematic diagram of a passive avoidance experiment.

[0023] Figure 4 is a graph showing the passive avoidance ability (sec) for the passive avoidance experiment of the normal group, the induced group, the positive control group 1, the positive control group 2, the comparative example 1 group, and the example 1 group.

[0024] Figure 5 is a schematic diagram of the Y-maze experiment.

[0025] Figure 6 is an image showing the movement tendency of mice in the Y-maze experiment of the normal group, the induced group, the positive control group 1, the positive control group 2, the comparative example 1 group, and the example 1 group.

[0026] Figure 7 is a graph quantitatively showing the movement trend of Figure 6.

[0027] Figure 8 is a graph evaluating spatial perception ability for a substitute experiment of the normal group, the induced group, the positive control group 1, the positive control group 2, the comparative example 1 group, and the example 1 group.

[0028] The present invention relates to a composition capable of improving cognitive function, comprising a phospholipid mixture, a neutral lipid, and a spinolipid.

[0029]

[0030] Hereinafter, the present invention will be described in detail.

[0031] The cognitive function improvement composition of the present invention comprises a phospholipid mixture, a neutral lipid, a spinolipid, and an oil.

[0032] The phospholipid mixture of the present invention comprises phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, glycosphingolipids, and gangliosides, and exhibits a superior effect in improving cognitive function compared to when only phosphatidylserine and phosphatidylethanolamine are used.

[0033] In particular, the phospholipid mixture comprises 80 to 95 parts by weight, preferably 85 to 90 parts by weight, of phosphatidylethanolamine, based on 100 parts by weight of phosphatidylcholine; 30 to 50 parts by weight, preferably 35 to 45 parts by weight, of phosphatidylserine; 20 to 40 parts by weight, preferably 25 to 35 parts by weight, of phosphatidylinositol; 50 to 80 parts by weight, preferably 60 to 70 parts by weight, of sphingomyelin; 80 to 95 parts by weight, preferably 85 to 92 parts by weight, of glycosphingolipids; And gangliosides, 5 to 20 parts by weight, preferably 10 to 15 parts by weight, and if the content is outside the above range, cognitive function may not be improved.

[0034] In addition, the neutral lipid of the present invention, when used together with a phospholipid mixture, exhibits a cognitive function improvement effect that is 2 to 5 times better than when the phospholipid mixture alone or the neutral lipid alone is used.

[0035] These neutral lipids are used in an amount of 40 to 70 parts by weight, preferably 50 to 60 parts by weight, and more preferably 53 to 56 parts by weight, per 100 parts by weight of the phospholipid mixture. If the content of the neutral lipid is below the lower limit, the cognitive function improvement effect may be minimal, and if it exceeds the upper limit, the cognitive function improvement effect may not be enhanced and the sensory properties may be reduced.

[0036] The neutral lipid comprises at least one selected from the group consisting of EPA and DHA, and may further comprise oleic acid and tocopherol. The oleic acid is contained in an amount of 20 to 100 parts by weight, preferably 25 to 60 parts by weight, based on 100 parts by weight of the at least one substance selected from the group consisting of EPA and DHA; the tocopherol is contained in an amount of 0.5 to 10 parts by weight, preferably 2 to 5 parts by weight, based on 100 parts by weight of the at least one substance selected from the group consisting of EPA and DHA. If the content of oleic acid and tocopherol exceeds the above range, the cognitive function improvement effect may not be enhanced.

[0037] The above EPA contains 800 to 1200 mg of EPA (ethyl ester preparation, EE), preferably 900 to 1100 mg; and 800 to 1200 mg of EPA (free fatty acid preparation, FFA), preferably 850 to 1100 mg; and the above DHA contains 300 to 1200 mg of DHA (ethyl ester preparation, EE), preferably 350 to 900 mg; and 300 to 1200 mg of DHA (free fatty acid preparation, FFA), preferably 320 to 1000 mg.

[0038] The above tocopherol is not particularly limited as long as it is used for anti-oxidation.

[0039] In addition, the sphingolipide of the present invention has a further enhanced cognitive function improvement effect when used together with a phospholipid mixture and neutral lipid, and is used in an amount of 20 to 50 parts by weight, preferably 30 to 40 parts by weight, per 100 parts by weight of the phospholipid mixture.

[0040] If the content of spinolipids is below the lower limit, the cognitive function improvement effect may not be excellent, and if it exceeds the upper limit, the cognitive function improvement effect may actually be reduced.

[0041] In addition, the above-mentioned oils may include fish oil composed of medium chain triglyceride (MCT) oil; one or more vegetable oils selected from the group consisting of perilla oil, evening primrose oil, black currant oil, borage oil, hemp seed oil, cottonseed oil, safflower oil, soybean oil, and sunflower oil; or a mixed oil thereof.

[0042] The above medium chain triglyceride (MCT) oil is a main component of refined processed oil, and is manufactured using C6 to C10 fatty acids contained in coconut oil or palm oil as the main raw material among fatty acid triacylglycerins having 10 or fewer carbon atoms. MCT oil is produced by esterifying the above fatty acids and glycerin, and is characterized by a fatty acid composition of C6 of 2 wt% or less, C8 of 35 to 85 wt%, C10 of 15 to 45 wt%, and C12 of 3 wt% or less. In addition, some linolenic acid (C18:2) of 4 to 6 wt% and succinic acid of 12 to 16 wt% are added to adjust the physical properties. Because it is a triacylglycerin composed solely of saturated fatty acids, it has superior oxidative stability compared to triacylglycerins containing unsaturated fatty acids such as soybean oil and olive oil, and has low viscosity, low solidification point, low surface tension, good extensibility, and excellent solubility and lubrication properties. In addition, MCT oil has several times higher solubility in many substances such as medicines, vitamins, amino acids, and coloring agents than soybean oil, one of the representative vegetable oils, and its excellent digestibility and absorbability can improve bioavailability. Because it is hydrolyzable and absorbable in the human body much faster than long-chain triglycerides (LCTs), which are conventional fats and oils, it is widely used as a therapeutic food for various malabsorption syndromes, such as cases where the nutrient absorption surface is reduced due to intestinal disease or surgery, and cases where lipase and bile acid secretion is insufficient due to pancreatic disease or liver disease.

[0043] The above-mentioned evening primrose oil, blackcurrant oil, borage oil, and hemp seed oil contain γ-linolenic acid, and the above-mentioned cottonseed oil, safflower oil, and soybean oil contain linolenic acid.

[0044] The above-mentioned oil is used in an amount of 130 to 160 parts by weight, preferably 145 to 155 parts by weight, per 100 parts by weight of the phospholipid mixture. If the content of the oil exceeds the above range, the cognitive function improvement effect is not affected.

[0045] The cognitive function improvement composition of the present invention contains 2 to 5 g / 100 g of lactose, preferably 3.0 to 3.5 g / 100 g; and 6 to 8 g / 100 g of ashes, preferably 6.5 to 7.5 g / 100 g. In addition, the pH of the cognitive function improvement composition is 6.3-6.7.

[0046]

[0047] In addition, the present invention can provide a food composition for improving brain function or a pharmaceutical composition for preventing and improving Alzheimer's disease, including the cognitive function composition.

[0048] The pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable auxiliary agents in addition to the above-mentioned effective ingredient, and the auxiliary agents can include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, glidants, or flavoring agents.

[0049] The above pharmaceutical composition may be preferably formulated as a pharmaceutical composition by additionally including one or more pharmaceutically acceptable carriers in addition to the above-described effective ingredients for administration.

[0050] The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an orally acceptable, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants, and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose, or beta-lactose, natural and synthetic gums such as corn sweetener, acacia, tracheacanth, or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, etc.

[0051] In the composition to be formulated as a liquid solution, acceptable pharmaceutical carriers include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components. If necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

[0052] Furthermore, it can be preferably formulated according to each disease or ingredient using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, as an appropriate method in the relevant field.

[0053] The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., and oral administration is preferred.

[0054] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. A skilled physician can easily determine and prescribe a dosage effective for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g / kg.

[0055] The pharmaceutical composition of the present invention can be manufactured in a unit dose form or can be manufactured by inserting it into a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granules, tablet or capsule, and may additionally include a dispersing agent or stabilizer.

[0056] In addition, the present invention provides a food composition for improving brain function containing a phospholipid mixture, neutral lipid, spinolipid, and fat as effective ingredients.

[0057] The food composition according to the present invention can be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements, and the like.

[0058] The food composition of the present invention may include, as active ingredients, a mixture of phospholipids, neutral lipids, spinolipids, and fats, as well as components commonly added during food manufacturing, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the carbohydrates described above include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavoring agents [thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured into a drink or beverage, in addition to the phospholipid mixture, neutral lipid, spinolipid, and fat of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included.

[0059] The present invention provides a health functional food comprising a food composition for improving brain function, which comprises the phospholipid mixture, neutral lipid, sphingolipid, and fat as active ingredients. A health functional food is a food prepared by adding a phospholipid mixture, neutral lipid, sphingolipid, and fat to food materials such as beverages, teas, spices, gums, and confectionery, or by manufacturing it in the form of capsules, powders, suspensions, etc., and which, when consumed, brings about a specific health effect. However, unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of phospholipid mixture, neutral lipid, spinolipid and oil added to such health functional foods cannot be uniformly regulated as it varies depending on the type of target health functional foods, but may be added within a range that does not damage the original taste of the food, and is usually in the range of 0.01 to 50 wt%, preferably 0.1 to 20 wt%, with respect to the target food. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, they may be added in the range of usually 0.1 to 100 wt%, preferably 0.5 to 80 wt%. In one specific example, the health functional food of the present invention may be in the form of pills, tablets, capsules or beverages.

[0060] The present invention also provides the use of a phospholipid mixture, neutral lipids, sphingolipids, and fats for the manufacture of a medicine or food for improving brain function. As described above, the phospholipid mixture, neutral lipids, sphingolipids, and fats can be used for the purpose of improving brain function.

[0061] The present invention also provides a method for preventing and improving Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a mixture of phospholipids, neutral lipids, sphingolipids and maintenance.

[0062]

[0063] In addition, the present invention provides a method for treating cognitive dysfunction, comprising the step of administering to a subject in need thereof an effective amount of a composition comprising a phospholipid mixture, a neutral lipid, a spinolipid, and a fat as active ingredients.

[0064] The above cognitive impairment may include at least one selected from the group consisting of dementia induced by chronic ultraviolet exposure, Alzheimer's disease, Huntington's disease, vascular dementia, ischemic stroke, traumatic brain injury, amnesia, Parkinson's disease, Pick disease, Creutzfeldt-Jakob disease, and mild cognitive impairment.

[0065] In addition, the related symptoms of the above cognitive dysfunction may include at least one selected from the group consisting of decreased learning ability, decreased memory, lethargy, decreased attention, depression, decreased hearing, analgesia, ahidrosis, and decreased discrimination.

[0066] The term "subject in need" as used herein refers to a mammal, preferably a human, that is the subject of treatment, observation or experimentation.

[0067] The term "effective amount" as used herein means the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue, animal, or human, as conceived by a researcher, veterinarian, physician, or other clinician, including an amount that induces alleviation of the symptoms of the disease or disorder. It will be apparent to those skilled in the art that the effective amount and frequency of administration of the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be readily determined by those skilled in the art, and can be adjusted according to various factors, including the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, the type of formulation, and the patient's age, weight, general health, sex, and diet, the time of administration, the route of administration, and the excretion rate of the composition, the treatment period, and concurrently used drugs. In the method for preventing, treating, or improving a disease of the present invention, in the case of adults, it is preferable to administer the phospholipid mixture and the neutral lipid mixture once or several times a day at a dosage of 0.001 g / kg to 10 g / kg.

[0068] In the treatment method of the present invention, a composition comprising a phospholipid mixture, a neutral lipid, a sphingolipid and a fat as active ingredients can be administered in a conventional manner via oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes.

[0069] Hereinafter, preferred examples are presented to help understand the present invention, but the following examples are only illustrative of the present invention, and it is obvious to those skilled in the art that various changes and modifications are possible within the scope and technical idea of ​​the present invention, and it is also natural that such changes and modifications fall within the scope of the appended patent claims.

[0070] Example 1.

[0071] phospholipid mixture

[0072] A phospholipid mixture was prepared by including 86.36 parts by weight of phosphatidylethanolamine, 43.18 parts by weight of phosphatidylserine, 27.27 parts by weight of phosphatidylinositol, 68.18 parts by weight of sphingomyelin, 90.91 parts by weight of glycosphingolipids, and 10 parts by weight of gangliosides, per 100 parts by weight of phosphatidylcholine.

[0073] neutral lipids

[0074] A neutral lipid mixture was prepared by including 40 parts by weight of oleic acid and 4 parts by weight of tocopherol per 100 parts by weight of EPA.

[0075] The above EPA contains 960 mg of EPA (ethyl ester preparation, EE) and 880 mg of EPA (free fatty acid preparation, FFA), and the above DHA contains 380 mg of DHA (ethyl ester preparation, EE) and 340 mg of DHA (free fatty acid preparation, FFA).

[0076] Cognitive function improvement composition

[0077] A composition for improving cognitive function was obtained by containing 54.23 parts by weight of neutral lipid, 37.96 parts by weight of spinolipid, and 151.84 parts by weight of medium-chain triglyceride oil for 100 parts by weight of the above phospholipid mixture.

[0078]

[0079] <Example Ⅰ>In vitro

[0080] Control group 1 used a complex containing 100 parts by weight of phosphatidylcholine and 43 parts by weight of phosphatidylserine, and control group 2 used a synthetic drug, choline alfoscerate (La-GPC).

[0081] Test Example 1. Measurement of nerve cell proliferation effect

[0082] The cognitive function improvement compositions of Example 1, Control Group 1, and Control Group 2 were treated at concentrations of 0.01, 0.1, 1, and 10 mg / ml to mouse hippocampal cells (HT-22 cells), cultured for 24 hours, and measured for cell viability to evaluate the effect on cell proliferation. The hippocampal cells (HT-22 cells) are a cell line derived from the mouse hippocampus and are one of the models that can evaluate the effect of test substances on oxidative damage caused by degenerative brain diseases. The functionality for improving cognitive function was evaluated through the HT-22 cell model.

[0083] HT-22 cell line, a mouse hippocampal cell line, was cultured in a cell incubator maintained at 37°C with 5% CO2 using DMEM medium containing 10% FBS at 1%. The cultured cell line was cultured at 1X10 4 The samples were cultured in a 96-well plate at a concentration of , and after 24 hours, when the cells attached, the samples were treated at various concentrations and cultured for 24 hours to observe cell proliferation.

[0084] The survival rate of HT22 neurons cultured under various concentration conditions was measured using the MTS assay. Specifically, after treating with MTS solution, the cells were cultured at 37°C for 3 hours, dissolved in DMSO, and the absorbance change was measured at 540 nm (MTS: 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)).

[0085] The cell viability for each sample was expressed as a percentage to evaluate the neuronal proliferation effect of the sample.

[0086] Figure 1 is a graph showing the cell survival rate of hippocampal cells (HT-22 cells) when treated with a cognitive function improvement composition (a) manufactured according to Example 1 of the present invention, a cognitive function improvement composition of control group 1 (b), and a cognitive function improvement composition of control group 2 (c).

[0087] As shown in FIGS. 1A to 1C, the cognitive function improvement compositions of Example 1, Control Group 1, and Control Group 2 all showed no cytotoxicity, and it was confirmed that the cognitive function improvement composition prepared according to Example 1 showed a superior neuronal cell proliferation effect compared to Control Group 1 and Control Group 2.

[0088] The cognitive function improvement composition manufactured according to Example 1 showed a significant neuronal proliferation effect at a concentration of 0.01 ug / ml or more, and it was confirmed that the neuronal proliferation effect was significantly observed only at a concentration of 0.1 ug / ml or more in Control Group 1 and only at a concentration of 0.01 ug / ml in Control Group 2.

[0089]

[0090] Test Example 2. Measurement of cytoprotective effect against β-amyloid

[0091] Beta-amyloid protein is a 36-43 amino acid peptide that is the main component of amyloid plaques found in the brains of Alzheimer's patients and is critically involved in the pathogenesis of Alzheimer's disease. After beta-amyloid precursor protein is synthesized in normal cells, it is degraded by β-secretase or γ-secretase to produce Aβ1-42 or Aβ1-43 with an elongated C-terminus. These aggregate and accumulate in the form of insoluble amyloid fibers, causing damage and cell death in brain neurons. Therefore, in vitro, Aβ1-42 treatment was performed on samples to induce a cognitive decline cell model, and the protective effect on neurons was confirmed.

[0092] HT-22 cells 1X10 4After culturing the cells in a 96-well plate for 24 hours at a concentration of 10 mM, stable cell attachment was confirmed. The samples were treated with various concentrations and cultured for 24 hours. After 24 hours, 10 mM beta-amyloid was treated, the degree of cell death was observed, and cell viability was evaluated using the MTS method.

[0093] In order to establish a cognitive decline cell model using HT-22 cells, the research conditions were examined by treating them with various concentrations of beta-amyloid. When treated with 5, 10, and 15 uM of beta-amyloid, the cell viability was 87%, 80%, and 76%, respectively, and it was confirmed that significant cell death occurred at all three concentrations. Therefore, the cognitive decline cell model in this study was treated with 10 uM of beta-amyloid, which is a concentration that is approximately 20% reduced compared to normal cells, and the neuroprotective effect of the sample was examined.

[0094] Figure 2 is a graph showing cell viability after treatment with 10 uM of beta-amyloid, the cognitive function improvement composition manufactured according to each Example 1, and the cognitive function improvement compositions of Control Group 1 and Control Group 2. The induction group is the group treated with 10 uM of beta-amyloid.

[0095] As illustrated in Fig. 2, the cognitive function improvement compositions of Example 1, Control Group 1, and Control Group 2 all exhibited cytoprotective effects against beta-amyloid. In particular, the cognitive function improvement composition manufactured according to Example 1 exhibited a more excellent cytoprotective effect against beta-amyloid than Control Groups 1 and 2, and thus is judged to be a functional material that can aid in neuroprotection.

[0096]

[0097] <Example II>In vivo

[0098] Among the samples that showed neuronal proliferation and neuronal protection effects in the above in vitro study, the compositions of Example 1 and Control Group 1 were selected based on their excellent effects, considering the unit price of raw materials and the manufacturing process, and the cognitive function improvement effect was evaluated using an in vivo cognitive function decline model. The cognitive function improvement effect was performed through behavioral evaluation and mechanism study, and the behavioral evaluation was performed using the passive avoidance test and the spatial memory test (Y-maze test).

[0099] animal testing

[0100] C57BL / 6J mice (8 weeks old, male) were purchased and raised in an animal breeding facility and used to evaluate cognitive function improvement. The experimental animals were maintained under the following breeding environmental conditions: temperature 22±3℃, relative humidity 30±10%, lighting time 12 h (08:00-20:00), and illumination intensity 150-300 Lux. No more than 5 animals were raised in polycarbonate cages. Solid feed for experimental animals (Orient) was used, and pre-filtered tap water was used, and the experiment was conducted under ad libitum conditions.

[0101] The cognitive function effects were evaluated by administering the sample at a high concentration for a short period of time (500 mg / kg, once daily for 7 days) to each group of 8 mice. The mouse administration dose was calculated based on the toxic concentration of choline alfoscerate, and the individual administration dose was administered orally based on the administration concentration according to the mouse body weight.

[0102] -6 groups of mice-

[0103] Normal group: 7 days of saline solution administration

[0104] Induction group: 7 days of saline administration followed by Scopolamine administration_1 mg / kg / day

[0105] Positive control group 1: Donepezil administered for 7 days (8 mg / kg / day) followed by Scopolamine administration (1 mg / kg / day)

[0106] Comparative Example 1 Group: Administered the composition of Control Group 1 for 7 days (500 mg / kg / day) followed by Scopolamine administration (1 mg / kg / day)

[0107] Example 1 group: 7 days of administration of the composition of Example 1 (500 mg / kg / day) followed by administration of Scopolamine (1 mg / kg / day)

[0108] Positive control group 2: 7-day administration of the composition of control group 2 (500 mg / kg / day) followed by administration of Scopolamine (1 mg / kg / day)

[0109]

[0110] Test Example 3. Passive Avoidance Experiment

[0111] The passive avoidance experiment is a research method that utilizes the characteristic of mice to prefer dark environments, and is used to enhance memory and assess explicit memory. The experiment is conducted using equipment consisting of two acrylic boxes, one dark and one bright, with a door between which the mouse can move. The floor of the acrylic box is lined with a stainless steel bar, to which an electric stimulus can be applied according to the experimenter's preferences. Before the experiment, the mouse is acclimated to the white space. After 30 seconds, the door is opened and the mouse moves to the dark space. After a maximum of 180 seconds, the door is closed and a 0.5 mA electric stimulus is delivered once for 2 seconds. After 10 seconds of the electric stimulus, the mouse is removed, calmed, and returned to its cage (Figure 3).

[0112] During the oral administration period, passive avoidance training was performed daily. On the 8th day of the experiment, 1 mg / kg of scopolamine was administered orally to decrease cognitive function, and this experiment was conducted (Fig. 3). This experiment was performed in the same manner as passive avoidance training, and the mice were measured to remember the electrical stimulation in the dark space, suppress their natural tendency to the dark space, and remain in the bright space within 300 seconds. If there was no movement after that, the experiment was stopped.

[0113] Figure 4 is a graph showing the passive avoidance ability (sec) for the passive avoidance experiment of the normal group, the induced group, the positive control group 1, the positive control group 2, the comparative example 1 group, and the example 1 group.

[0114] There were no significant changes in body weight in any group of mice during oral administration and passive avoidance training. During passive avoidance training, all groups showed a tendency to remember the electrical stimulation and increase their latency time, confirming that training in the behavioral test was appropriate.

[0115] As shown in Fig. 4, it was confirmed that the latency time was significantly reduced in the induced group compared to the normal group, and it was confirmed that the latency time increased in all of the positive control group 1, positive control group 2, comparative example 1 group, and example 1 group, but it was confirmed that the latency time increased more in example 1 group than in the other groups.

[0116] Therefore, it was confirmed that the cognitive function improvement composition manufactured according to Example 1 was effective in improving cognitive function, and it was confirmed that this was at a similar level to that of the synthetic drug control group 2 (positive control group 2).

[0117]

[0118] Test Example 4. Y-Maze Model Experiment

[0119] The Y-maze experiment is a study comparing the spatial memory of rodents. It is an evaluation method that compares the ability to find a new path by remembering the path taken previously. The purpose of this study was to evaluate the effect of sample intake on improving spatial memory ability.

[0120] Mice generally tend to explore new branches of a maze rather than continue on previously entered branches. Mice with impaired cognitive function exhibit a reduced tendency to explore new branches, while those with enhanced cognitive function exhibit increased ability to explore new branches. Therefore, we evaluated the ability of functional candidate materials to enhance spatial cognition through mouse behavioral analysis.

[0121] The mouse was placed at the end of one of three identical branches (8 cm wide, 30 cm long, and 14 cm high) arranged at a 120° angle and allowed to freely roam the maze for 5 minutes. The number of times and order in which it entered each branch were recorded. The path and distance traveled during the 5 minutes were checked to observe behavioral changes due to cognitive decline (Fig. 5).

[0122] Figure 6 is an image showing the movement tendency of mice in the Y-maze experiment of the normal group, the induced group, the positive control group 1, the positive control group 2, the comparative example 1 group, and the example 1 group; Figure 7 is a graph quantitatively showing the movement tendency of Figure 6.

[0123] As illustrated in Figure 6, the movement trends visualized in the images from the recording of mouse movement paths in the Y-maze experiment confirmed that the movement of mice and behavioral changes such as entering new branches were reduced in the induction group. In addition, it was confirmed that the movements of Example 1 Group, Comparative Example 1 Group, Positive Control Group 1, and Positive Control Group 2 were at a similar level to the normal group.

[0124] In addition, as shown in Fig. 7, the total number of times the branch entered the moving trend was recorded and quantitatively expressed as a result, and it was confirmed that the positive control group 1, positive control group 2, comparative example 1 group, and example 1 group increased compared to the induction group, and that the example 1 group increased more significantly than the other groups.

[0125]

[0126] Test Example 5. Alternation Behavior Test

[0127] In the alternation behavior test, spatial perception was assessed by recording the number of consecutive entries into each branch. Mice generally tend to explore new arms of the maze rather than reenter previously entered branches. Therefore, the number of times they re-entered a previously entered branch was assessed as a decrease in memory.

[0128] Figure 8 is a graph evaluating spatial perception ability for a substitute experiment of the normal group, the induced group, the positive control group 1, the positive control group 2, the comparative example 1 group, and the example 1 group.

[0129] As shown in Figure 8, the number of sequential entries (alternative time) in each branch increased in all groups compared to the induction group, but it was confirmed that the number of sequential entries in group 1 increased more significantly than in the other groups.

[0130]

[0131] Below, a formulation example of a composition containing the powder of the present invention is described, but the present invention is not intended to be limited thereto, but is merely intended to be described specifically.

[0132] Preparation Example 1. Preparation of a powder

[0133] 500 mg of the composition obtained in Example 1

[0134] 100 mg of lactose

[0135] 10 mg of talc

[0136] The above ingredients are mixed and filled into a sealed bag to prepare a powder.

[0137]

[0138] Preparation Example 2. Preparation of tablets

[0139] 300 mg of the composition obtained in Example 1

[0140] 100 mg of corn starch

[0141] 100 mg of lactose

[0142] Magnesium stearate 2 mg

[0143] After mixing the above ingredients, tablets are manufactured by pressing them according to the usual tablet manufacturing method.

[0144]

[0145] Preparation Example 3. Preparation of capsules

[0146] 200 mg of the composition obtained in Example 1

[0147] 3 mg of crystalline cellulose

[0148] 14.8 mg of lactose

[0149] Magnesium stearate 0.2 mg

[0150] The above ingredients are mixed according to the conventional capsule manufacturing method and filled into a gelatin capsule to manufacture a capsule.

[0151]

[0152] Preparation Example 4. Preparation of Injectable

[0153] 600 mg of the composition obtained in Example 1

[0154] Mannitol 180 mg

[0155] 2974 mg of sterile distilled water for injection

[0156] Na2HPO 4, 12H2O 26 mg

[0157] It is manufactured with the above ingredient content per ampoule according to the manufacturing method of a conventional injection.

[0158]

[0159] Preparation Example 5. Preparation of liquid preparation

[0160] 4 g of the composition obtained in Example 1

[0161] 10 g of isoflavonoids

[0162] 5 g of mannitol

[0163] Appropriate amount of purified water

[0164] According to the usual method of manufacturing a liquid, each ingredient is added to purified water and dissolved, an appropriate amount of lemon flavor is added, the above ingredients are mixed, purified water is added, and the total amount is adjusted to 100 g, then filled into a brown bottle and sterilized to manufacture a liquid.

[0165]

[0166] Preparation Example 6. Preparation of granules

[0167] 1,000 mg of the composition obtained in Example 1

[0168] Vitamin mixture appropriate amount

[0169] Vitamin A acetate 70 μg

[0170] Vitamin E 1.0 mg

[0171] Vitamin B1 0.13 mg

[0172] Vitamin B2 0.15 mg

[0173] Vitamin B6 0.5 mg

[0174] Vitamin B12 0.2 μg

[0175] Vitamin C 10 mg

[0176] 10 μg of biotin

[0177] 1.7 mg of nicotinamide

[0178] 50 μg of folic acid

[0179] Calcium pantothenate 0.5 mg

[0180] Appropriate amount of mineral mixture

[0181] 1.75 mg of ferrous sulfate

[0182] 0.82 mg of zinc oxide

[0183] Magnesium carbonate 25.3 mg

[0184] 15 mg of monobasic potassium phosphate

[0185] 55 mg of dibasic calcium phosphate

[0186] 90 mg of potassium citrate

[0187] 100 mg of calcium carbonate

[0188] Magnesium chloride 24.8 mg

[0189] The composition ratio of the above vitamin and mineral mixture is a preferred example of mixing ingredients relatively suitable for granules, but the mixing ratio may be arbitrarily modified, and the above ingredients may be mixed according to a conventional granule manufacturing method, and then granules may be manufactured and used in the manufacture of a health functional food composition according to a conventional method.

[0190]

[0191] Preparation Example 7. Manufacturing of functional beverages

[0192] 1,000 mg of the composition obtained in Example 1

[0193] 1,000 mg of citric acid

[0194] 100 g of oligosaccharide

[0195] 2 g of plum concentrate

[0196] 1 g of taurine

[0197] Add purified water to make a total of 900 mL

[0198] The above ingredients are mixed according to a conventional health beverage manufacturing method, then stirred and heated at 85°C for about 1 hour, the resulting solution is filtered, placed in a sterilized 2 L container, sealed and sterilized, and then stored in a refrigerator, and then used to manufacture the functional beverage composition of the present invention.

[0199] The above composition ratio is a preferred example of a mixture of ingredients relatively suitable for a preferred beverage, but the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demand country, and intended use.

[0200] The cognitive function improvement composition of the present invention is effective in improving cognitive function, and therefore can be used not only as a food composition for improving brain function but also as a pharmaceutical composition for preventing and improving Alzheimer's disease.

Claims

1. A mixture of phospholipids comprising phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyeline, glycosphingolipids and gangliosides; Neutral lipids comprising at least one selected from the group consisting of EPA and DHA; sphingolipides; and A composition for improving cognitive function, characterized by including a maintenance; 2. A composition for improving cognitive function, characterized in that in claim 1, the composition contains 40 to 70 parts by weight of neutral lipid, 20 to 50 parts by weight of spinolipid, and 130 to 160 parts by weight of fat per 100 parts by weight of the phospholipid mixture.

3. A composition for improving cognitive function, characterized in that in claim 1, the phospholipid mixture comprises 80 to 95 parts by weight of phosphatidylethanolamine, 30 to 50 parts by weight of phosphatidylserine, 20 to 40 parts by weight of phosphatidylinositol, 50 to 80 parts by weight of sphingomyeline, 80 to 95 parts by weight of glycosphingolipids, and 5 to 20 parts by weight of gangliosides, per 100 parts by weight of phosphatidylcholine.

4. A food composition for improving brain function, comprising a cognitive function composition according to any one of claims 1 to 3.

5. A pharmaceutical composition for preventing and improving Alzheimer's disease, comprising a cognitive function composition according to any one of claims 1 to 3 as an active ingredient.

6. A method for treating cognitive dysfunction, comprising the step of administering to a subject in need of a cognitive function composition according to any one of claims 1 to 3.

7. A method for treating cognitive dysfunction in claim 12, characterized in that the cognitive dysfunction is at least one selected from the group consisting of dementia induced by chronic ultraviolet exposure, Alzheimer's disease, Huntington's disease, vascular dementia, ischemic stroke, traumatic brain injury, amnesia, Parkinson's disease, Pick disease, Creutzfeldt-Jakob disease, and mild cognitive dysfunction.

8. A method for treating cognitive dysfunction in accordance with claim 12, characterized in that the related symptoms of cognitive dysfunction are at least one selected from the group consisting of decreased learning ability, decreased memory, lethargy, decreased attention, depression, decreased hearing, analgesia, ahidrosis, and decreased discrimination.

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