KR102147894B1 - Compositon for Cognitive Function Activity Effect comprising phospholipid mixture, vitamin mixture, neutral lipid mixture - Google Patents

Abstract

The present invention relates to a composition for improving a cognitive function. The composition includes: a phospholipid mixture containing phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidic acid; a neutral lipid mixture containing EPA, DHA and tocopherol; a vitamin mixture containing vitamin E, vitamin C, vitamin D and vitamin B12; and oil and fats. The composition shows an activity of removing active oxygen species, an antioxidative activity against DNA damages, an activity of inhibiting ACE (angiotensin converting enzyme), an activity of inhibiting AChE (acetylcholine esterase), and an activity of inhibiting production of NO (nitric oxide).

Description

Cognitive function improvement composition comprising phospholipid mixture, vitamin mixture, and neutral lipid mixture {Compositon for Cognitive Function Activity Effect comprising phospholipid mixture, vitamin mixture, neutral lipid mixture}

The present invention provides a composition for improving cognitive function that can exhibit the removal of reactive oxygen species, antioxidant against DNA damage, ACE (Angiotensin Converting Enzyme) enzyme inhibitory activity, AChE (acetylcholine esterase) inhibitory activity, NO (Nitric oxide) production inhibitory activity. About.

The human brain is an important part that manages learning, memory, and movement, and neurons are connected to each other to form a huge neural network, which is the basis of the nervous system including learning and memory.

However, when reactive oxygen species are excessively produced and accumulated in the body due to various causes, they cause oxidative damage to cells, leading to cell degeneration and cell death. In normal cases, it is removed by superoxide dismutase (SOD), catalase, carotenoid, and glutathione, which are the elimination mechanisms in the body, but ROS remaining in the body induces degeneration and death of brain neurons, negatively affecting the formation of the correct neural network. And, in severe cases, causes degenerative brain diseases such as Parkinson's disease and Alzheimer's.

In addition, cognitive abilities such as learning and memory in the brain are expressed by neurotransmitters such as acetylcholine (ACh). ACh secreted from the nerve cell ends normally transmits signals through synaptic receptors for external stimuli such as learning, and is then decomposed into acetate and choline by acetycholinesterase (AChE) and partially reabsorbed. This process is called the cholinergic system, and most of Alzheimer's patients are known to have serious problems during this process. Decreased ACh impairs the brain's cognitive ability, preventing it from functioning normally.

Therefore, AChE inhibitors are widely used as treatments for Alzheimer's patients. In addition, hypertension is also a cerebral vascular disease that causes stroke and arteriosclerosis, and is induced by Angiotensin II. Angiotensin II is an enzyme inducing hypertension in which angiotensine degraded by renine is converted by angiotensin I converting enzyme (ACE). This angiotensin II increases blood pressure by decomposing bradykinin, an enzyme that constricts arterial blood vessels and expands blood vessels. In addition, stress can also cause high blood pressure.

Korean Patent Registration No. 0864380 Republic of Korea Patent Publication No. 2012-0131149

An object of the present invention is to improve cognitive function that can show the removal of reactive oxygen species, antioxidant against DNA damage, ACE (Angiotensin Converting Enzyme) enzyme inhibitory activity, AChE (acetylcholine esterase) inhibitory activity, NO (Nitric oxide) production inhibitory activity It is to provide a composition.

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

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

The composition for improving cognitive function of the present invention for achieving the above object includes phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol, phosphatidic acid, and saccharides. A mixture of phospholipids; Neutral lipid mixtures including EPA, DHA, oleic acid and tocopherol; Vitamin mixtures including vitamin E, vitamin C, vitamin D and vitamin B12; And maintenance; may include.

The composition may be contained in an amount of 0.1 to 10 parts by weight of a neutral lipid mixture, 0.1 to 5 parts by weight of a vitamin mixture, and 1 to 10 parts by weight of fats and oils based on 100 parts by weight of the phospholipid mixture.

The phospholipid mixture is phosphatidylcholine 10 to 50 parts by weight, phosphatidylethanolamine 20 to 50 parts by weight, phosphatidylinositol 5 to 30 parts by weight, phosphatidylinositol 5 to 30 parts by weight, based on 100 parts by weight of phosphatidylcholine. (phosphatidic acid) may contain 20 to 50 parts by weight and 10 to 30 parts by weight of saccharides.

The saccharides may be monosaccharides, disaccharides, oligosaccharides or polysaccharides.

The neutral lipid mixture may contain 25 to 100 parts by weight of DHA, 20 to 100 parts by weight of oleic acid, and 0.5 to 10 parts by weight of tocopherol based on 100 parts by weight of EPA.

The vitamin mixture may be a mixture of vitamin E, vitamin C, vitamin D, and vitamin B12 in a weight ratio of 1: 0.5-2: 0.1-1: 1-10.

The vitamin E is D-alpha-tocopherol (D-α-Tocopherol) based on 100 parts by weight of DL-alpha-tocopherol (DL-α-Tocopherol) 10 to 80 parts by weight, D-alpha-tocopheryl acid succinic acid (D- α-Tocopheryl Acid Succinate) 5 to 70 parts by weight, D-alpha-Tocopheryl Acetate 10 to 70 parts by weight and DL-alpha-Tocopheryl Acetate 5 to It may be a mixture containing 70 parts by weight.

The fats and oils are fish oil consisting of medium chain triglyceride (MCT) oil; One or more vegetable oils selected from the group consisting of perilla oil, evening primrose oil, blackcurrant oil, borage oil, hemp seed oil, cottonseed oil, safflower oil, soybean oil and sunflower oil; Or it may be a mixture of these.

In addition, the food composition for improving brain function of the present invention for achieving the above-described other object may include the cognitive function composition.

In addition, the pharmaceutical composition for preventing and improving Alzheimer's according to the present invention for achieving the another object described above may include the cognitive function composition.

Cognitive function improvement composition of the present invention removes reactive oxygen species, antioxidant against DNA damage, ACE (Angiotensin Converting Enzyme) enzyme inhibitory activity, AChE (acetylcholine esterase) inhibitory activity, NO (Nitric oxide) production inhibitory activity. It can be used not only to improve, but also to pharmaceutical compositions for preventing and improving Alzheimer's.

The present invention provides a composition for improving cognitive function that can exhibit the removal of reactive oxygen species, antioxidant against DNA damage, ACE (Angiotensin Converting Enzyme) enzyme inhibitory activity, AChE (acetylcholine esterase) inhibitory activity, NO (Nitric oxide) production inhibitory activity. About.

Hereinafter, the present invention will be described in detail.

The composition for improving cognitive function of the present invention includes a phospholipid mixture, a neutral lipid mixture, a vitamin mixture, and a fat or oil.

Phospholipid mixture of the present invention is phosphatidylserine (phosphatidylcholine), phosphatidylcholine (phosphatidylcholine), phosphatidylethanolamine (phosphatidylethanolamine), phosphatidylinositol (phosphtidylinositol), phosphatidylinositol (phosphtidylinositol), phosphatidic acid (phosphatidylcholine) and phosphatidic acid (phosphatidylcholine) and saccharides containing Compared to the case of using only phosphatidylethanolamine, it has an excellent effect on improving cognitive function.

In particular, the phospholipid mixture may include 10 to 50 parts by weight of phosphatidylcholine, preferably 10 to 40 parts by weight, based on 100 parts by weight of phosphatidylcholine; Phosphatidylethanolamine 20 to 50 parts by weight, preferably 30 to 40 parts by weight; 5 to 30 parts by weight, preferably 10 to 20 parts by weight of phosphatidylinositol; Phosphatidic acid (phosphatidic acid) 20 to 50 parts by weight, preferably 30 to 40 parts by weight; And 10 to 30 parts by weight, preferably 20 to 25 parts by weight of saccharide, and if the content is outside the above range, the antioxidant and ACE enzyme inhibitory activity against DNA damage may be significantly reduced.

Although it does not specifically limit as said saccharide, Preferably, monosaccharide, disaccharide, oligosaccharide, or polysaccharide is mentioned.

In addition, the neutral lipid mixture of the present invention is used in combination with a phospholipid mixture, which is 2 to 5 times superior to the case of using a phospholipid mixture alone or a neutral lipid mixture alone. Antioxidant, ACE enzyme inhibitory activity, AChE inhibitory activity and It shows NO production inhibitory activity.

The neutral lipid mixture is used in an amount of 0.1 to 10 parts by weight, preferably 0.1 to 3 parts by weight, and more preferably 0.1 to 1 part by weight, based on 100 parts by weight of the phospholipid mixture. Even if a small amount of a neutral lipid mixture is used, the desired effect can be remarkably improved, and if the content of the neutral lipid mixture is less than the above lower limit, the desired effect may be reduced, and if it exceeds the upper limit, the effect of improving cognitive function may be insignificant. have.

The neutral lipid mixture is DHA (docosahexaenoic acid) 25 to 100 parts by weight, preferably 35 to 80 parts by weight based on 100 parts by weight of EPA (eicosapentaenoic acid); 20 to 100 parts by weight of oleic acid, preferably 25 to 60 parts by weight; And 0.5 to 10 parts by weight of tocopherol, preferably 2 to 5 parts by weight, and if the content is outside the above range, the ACE enzyme inhibitory activity, AChE inhibitory activity, and NO production inhibitory activity may be significantly reduced.

In particular, the EPA is EPA (ethyl ester formulation, EE) 800 to 1200 mg, preferably 900 to 1100 mg; And EPA (free fatty acid preparation, FFA) 800 to 1200 mg, preferably 850 to 1100 mg, and the DHA contains DHA (ethyl ester preparation, EE) 300 to 1200 mg, preferably 350 to 900 mg; And DHA (free fatty acid preparation, FFA) 300 to 1200 mg, preferably 320 to 1000 mg.

The tocopherol is not particularly limited as long as it is used to prevent oxidation.

In addition, the vitamin mixture of the present invention further enhances the antioxidant against DNA damage, ACE enzyme inhibitory activity, AChE inhibitory activity, NO production inhibitory activity when used with a phospholipid mixture and a neutral lipid mixture. In the case of using vitamins, antioxidant activity against DNA damage, ACE enzyme inhibitory activity, AChE inhibitory activity, and NO production inhibitory activity may not be improved.

This vitamin mixture is used in an amount of 0.1 to 5 parts by weight, preferably 0.5 to 3 parts by weight, based on 100 parts by weight of the phospholipid mixture. When the content of the vitamin mixture is less than the lower limit, the desired effect cannot be improved, and when the content of the vitamin mixture exceeds the upper limit, there is no effect on improving cognitive function.

The vitamin mixture includes vitamin E, vitamin C, vitamin D and vitamin B12, and preferably, tamin E, vitamin C, vitamin D, and vitamin B12 are in a weight ratio of 1: 0.5-2: 0.1-1: 1-10. They are mixed, and more preferably, they are mixed in a weight ratio of 1:0.8-1.5:0.3-0.8:2-5. If the content of vitamin C, vitamin D, and vitamin B12 based on vitamin E is out of the above range, there is no effect on improving antioxidant activity and cognitive function.

Specifically, when vitamin E is used with other vitamins, antioxidant activity is further improved and effective for cognitive function, and DL-alpha-tocopherol (DL-α-) based on 100 parts by weight of D-alpha-tocopherol (D-α-Tocopherol). Tocopherol) 10 to 80 parts by weight, preferably 20 to 70 parts by weight; 5 to 70 parts by weight, preferably 20 to 50 parts by weight of D-alpha-Tocopheryl Acid Succinate; D-alpha-tocopheryl acetate (D-α-Tocopheryl Acetate) 10 to 70 parts by weight, preferably 25 to 55 parts by weight and DL-alpha-Tocopheryl Acetate (DL-α-Tocopheryl Acetate) 5 to 70 parts by weight , Preferably 10 to 40 parts by weight. When the content of vitamin E is outside the above range, the ACE enzyme inhibitory activity, AChE inhibitory activity, and NO production inhibitory activity may be significantly reduced.

When used with vitamin E and vitamin D, vitamin C further improves antioxidant activity, vitamin D may be effective in improving antioxidant activity and cognitive function when used with vitamin E, and vitamin B12 when used with vitamin E It is more effective in improving cognitive function.

In addition, the oil is fish oil consisting of medium chain triglyceride (MCT) oil; One or more vegetable oils selected from the group consisting of perilla oil, evening primrose oil, blackcurrant oil, borage oil, hemp seed oil, cottonseed oil, safflower oil, soybean oil and sunflower oil; Or a mixed oil and fat of these are mentioned.

The medium chain triglyceride (MCT) oil is a major component of refined processed fats and oils, fatty acid triacyl glycerins having 10 or less carbon atoms, and is prepared from C6 to C10 fatty acids contained in palm oil or palm oil as the main raw material. MCT oil is produced by esterifying the fatty acids and glycerin, and the fatty acid composition is composed of 2% by weight or less of C6, 35 to 85% by weight of C8, 15 to 45% by weight of C10, and 3% by weight of C12. It features. In addition, some linolenic acid (C18:2) 4-6% by weight and succinic acid 12-16% by weight are added to adjust the physical properties. As it is a triacyl glycerin consisting of only saturated fatty acids, it has excellent oxidation stability compared to triacyl glycerin having unsaturated fatty acids such as soybean oil and olive oil, and has low viscosity, low freezing point, low surface tension, good extensibility, and excellent solubility and lubrication. It has characteristics. In addition, MCT oil can improve bioavailability because its solubility in a number of substances such as medicines, vitamins, amino acids, coloring agents is several times higher than that of soybean oil, one of the representative vegetable oils, and has excellent digestion and absorption properties. Since the hydrolyzability and absorption in the human body is much faster than long-chain triglyceride (LCT), which is a common fat, the absorption area of nutrients is narrowed due to intestinal disease or surgery, due to pancreatic disease and liver disease. It is widely used as a treatment for various malabsorption syndromes, such as insufficient secretion of lipase and bile acids.

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

The fat or oil is used in an amount of 1 to 10 parts by weight, preferably 3 to 5 parts by weight, based on 100 parts by weight of the phospholipid mixture. If the content of fats and oils is less than the lower limit, antioxidant and ACE enzyme inhibitory activity against DNA damage may not be improved, and if the content of fat or oil is greater than the upper limit, the ACE enzyme inhibitory activity, AChE inhibitory activity, and NO production inhibitory activity may rather decrease. have.

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

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, lubricants. Agents or flavoring agents can be used.

For administration, the pharmaceutical composition may contain at least one pharmaceutically acceptable carrier in addition to the above-described active ingredient, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be 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 oral, 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 are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth 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, and the like.

In the composition formulated as a liquid solution, acceptable pharmaceutical carriers are sterilization and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and these One or more of the components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

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

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration. .

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, Usually, the skilled practitioner can readily determine and prescribe the dosage effective for the desired treatment or prophylaxis. 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.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the art. Or it can be made by incorporating it into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention provides a food composition for improving brain function containing a mixture of phospholipids and a mixture of neutral lipids as active ingredients.

The food composition according to the present invention may 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. Etc.

The food composition of the present invention may include not only a mixture of phospholipids and a mixture of neutral lipids as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Includes. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, 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 made of drinks and beverages, in addition to the phospholipid mixture and the neutral lipid mixture of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts are additionally included. I can make it.

The present invention provides a health functional food comprising a food composition for improving brain function comprising the phospholipid mixture and the neutral lipid mixture as an active ingredient. Health functional foods are foods prepared by adding a mixture of phospholipids and a mixture of neutral lipids to food materials such as beverages, teas, spices, gums, confectionery, etc., or prepared by encapsulation, powdering, or suspension. It means bringing about, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time 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 and neutral lipid mixture added in such health functional foods cannot be uniformly regulated depending on the type of the target health functional food, but can be added within the range that does not damage the original taste of the food. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides a use of a phospholipid mixture and a neutral lipid mixture for the manufacture of a medicine or food for improving brain function. As described above, the phospholipid mixture and the neutral lipid mixture may be used for improving brain function.

In addition, the present invention provides a method for the prevention and improvement of Alzheimer's comprising administering to a mammal an effective amount of a mixture of phospholipids and a mixture of neutral lipids.

The term "mammal" as used herein refers to a mammal that is the subject of treatment, observation or experimentation, and preferably refers to a human.

The term "effective amount" as used herein refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, Includes an amount that induces relief of symptoms of the disease or disorder. It is apparent to those skilled in the art that the effective amount and the number of administrations for the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and secretion rate of the composition, duration of treatment, and drugs used simultaneously. In the prophylaxis, treatment or improvement method of the present invention, in the case of adults, it is preferable to administer the phospholipid mixture and the neutral lipid mixture at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.

In the treatment method of the present invention, the composition comprising a phospholipid mixture and a neutral lipid mixture as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route. It can be administered in a conventional manner.

Hereinafter, a preferred embodiment is presented to aid the understanding of 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 spirit of the present invention, It is natural that such modifications and modifications fall within the appended claims.

Example 1.

Phospholipid mixture

37.5 parts by weight of phosphatidylcholine, 37.5 parts by weight of phosphatidylethanolamine, 12.5 parts by weight of phosphatidylinositol, 25.0 parts by weight of phosphatidylinositol, 37.5 parts by weight of phosphatidic acid, based on 100 parts by weight of phosphatidylcholine A phospholipid mixture was prepared including parts by weight.

Neutral lipid mixture

A neutral lipid mixture was prepared including 40 parts by weight of DHA, 40 parts by weight of oleic acid, and 4 parts by weight of tocopherol based on 100 parts by weight of EPA.

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

Vitamin mixture

Vitamin E, vitamin C, vitamin D, and vitamin B12 were mixed in a weight ratio of 1: 1.2: 0.5: 5 to prepare a vitamin mixture. At this time, the vitamin E is 40 parts by weight of DL-alpha-tocopherol, 40 parts by weight of D-alpha-tocopheryl acid succinic acid (D-α) based on 100 parts by weight of D-alpha-tocopherol (D-α-Tocopherol). -Tocopheryl Acid Succinate) 30 parts by weight, D-alpha-tocopheryl acetate (D-α-Tocopheryl Acetate) 50 parts by weight, and DL-alpha-tocopheryl acetate (DL-α-Tocopheryl Acetate) 40 parts by weight.

Cognitive function improvement composition

A composition for improving cognitive function was obtained by containing 0.1 parts by weight of a neutral lipid mixture, 0.6 parts by weight of a vitamin mixture, and 3.3 parts by weight of a medium chain triglyceride oil based on 100 parts by weight of the phospholipid mixture.

Example 2. Vegetable oil addition

In the same manner as in Example 1, but instead of 3.3 parts by weight of medium-chain triglyceride oil, 1.1 parts by weight of medium-chain triglyceride oil, 1.1 parts by weight of perilla oil, and 1.1 parts by weight of decoction oil were mixed to obtain a composition for improving cognitive function.

Comparative Example 1.

Phospholipid mixture

Phosphatidylserine (phosphatidylcholine) 100 parts by weight and phosphatidylethanolamine (phosphatidylethanolamine) 150 parts by weight were mixed to prepare a phospholipid mixture.

Neutral lipid mixture

A neutral lipid mixture was prepared by mixing 100 parts by weight of EPA and 40 parts by weight of DHA.

Vitamin mixture

Vitamin E, vitamin C, vitamin D, and vitamin B12 were mixed in a weight ratio of 1: 0.5: 0.1: 0.5 to prepare a vitamin mixture. At this time, the vitamin E is 40 parts by weight of DL-alpha-tocopherol, 40 parts by weight of D-alpha-tocopheryl acid succinic acid (D-α) based on 100 parts by weight of D-alpha-tocopherol (D-α-Tocopherol). -Tocopheryl Acid Succinate) 30 parts by weight, D-alpha-tocopheryl acetate (D-α-Tocopheryl Acetate) 50 parts by weight, and DL-alpha-tocopheryl acetate (DL-α-Tocopheryl Acetate) 40 parts by weight were included.

Cognitive function improvement composition

A composition for improving cognitive function was obtained by containing 0.1 parts by weight of a neutral lipid mixture, 0.6 parts by weight of a vitamin mixture, and 3.3 parts by weight of a medium chain triglyceride oil based on 100 parts by weight of the phospholipid mixture.

<Test Example>

Test Example 1. In vitro lipid peroxidation inhibition

In vitro lipid peroxidation inhibition: After mixing the compositions of Example 1 and Comparative Example 1 with linolenic acid emulsion, a solution of 0.8 mM H2O2 and 0.8 mM FeSO4 was reacted for 5 hours, then 0.4% TBA was added and at 95°C. After reacting for 2 hours, it was reacted at room temperature for 10 minutes. Then, 500 μl of a 15:1 ratio n-butanol:pyridine solution was added, centrifuged at 1,000 × g for 10 minutes, and the supernatant was measured for absorbance at 532 nm. The degree of lipid peroxidation was determined by the absorbance ratio before and after the addition of the sample. It is expressed in terms of% value. 0.1% vitamin E was used as a positive control.

division blank Vitamin E Example 1 Example 2 Comparative Example 1 Inhibition of lipid peroxidation (%) 100 60.4 65.1 63.0 98.1

As shown in Table 1 above, it was confirmed that the compositions prepared according to Examples 1 and 2 of the present invention had superior lipid peroxidation inhibitory effect compared to Comparative Example 1, which was similar to vitamin E used as a modal control. .

Test Example 2. Analysis of DNA damage by hydroxy radical

Genomic DNA was extracted from PC12 cells according to a slightly modified standard procedure (Sambrook J, Russell DW. Molecular cloning: a laboratory manual. New York, CSHL Press, 2001.). The oxidation of DNA exposed to hydroxyl radicals generated by the Fenton reaction was carried out according to a previously tested method (Milne L, Nicotera P, Orrenius S, Burkitt M. Effects of glutathione and chelating agents on copper-mediated DNA oxidation: pro. -oxidant and antioxidant properties of glutathione.Arch Bi°Chem Biophys 304, 102-109, 1993.). First, the compositions of Example 1 and Comparative Example 1, 200 μM FeSO4, 1 mM H2O2, and 50 μg/ml genomic DNA were added to 100 μl of a DNA solution. The reaction mixture was reacted at room temperature for 30 minutes, and then 10 mM EDTA was added to terminate the reaction. 20 μl of the reaction mixture containing 1 μg of DNA was electrophoresed on a 1% agarose gel at 100 V for 30 minutes. The gel was stained with 1 mg/ml ethidium bromide, washed with water, and observed with UV using a LAS3000 image analyzer (Fujifilm Life Science, Tokyo, Japan).

division blank ·OH Example 1 Example 2 Comparative Example 1 DNA oxidation (%) 100 71.5 207.1 224.1 148.5

Decomposition occurs when genomic DNA is exposed to the Hydroxyl radical produced by the Fenton reaction. In order to investigate the antioxidant effect of the compositions prepared according to Example 1, Example 2 and Comparative Example 1 on DNA damage, genomic DNA was isolated from PC12 cells and exposed to hydroxyl radicals generated by a fenton reaction.

As shown in Table 2 above, the compositions prepared according to Examples 1 and 2 of the present invention inhibited DNA degradation compared to Comparative Example 1, so that the compositions of Examples 1 and 2 significantly reduced DNA damage caused by hydroxyl radicals. It was confirmed that there is a decrease.

Test Example 3. Cell proliferation effect measurement

Cell culture: PC12 cells were cultured in DMEM medium containing 10% fetal bovine serum, 2 mM glutamine, and 100 μg/μl penicillin-streptomycin in an incubator maintained at 95% or higher humidity at 37° C. and 5% CO2.

MTT assay: Method of Hansen et al. (Hansen MB, Nielsen SE, Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Meth 119, 203-210, 1989.) According to the cytotoxicity of the Example for PC12 cells was measured using MTT (3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide).

division blank Example 1 Example 2 Comparative Example 1 Cell viability (%) 99.8 99.7 99.6 99.4

As shown in Table 3 above, it was confirmed that the compositions prepared according to Example 1, Example 2 and Comparative Example 1 of the present invention were not toxic as it was considered to have a high cell survival rate.

Test Example 4. ACE (Angiotensin I Converting Enzyme) Inhibitory Activity Measurement

ACE activity was obtained by extracting rabbit lung acetone powder in 0.1 M sodium borate buffer (pH 8.3) containing 0.3 M NaCl at a concentration of 1 g/10 ml (w/v) at 4°C for 2 hours, followed by centrifugation (4 ℃, 4,000 rpm, 40 minutes) to obtain an ACE coenzyme solution. Example 1, Comparative Example 1 and positive control (0.1% Captopril) 100 μl of 0.1 M sodium borate buffer (pH 8.3) and 50 μl of ACE coenzyme were added, and then pre-reacted at 37° C. for 5 minutes. , 50 μl of a substrate prepared by adding 25 mg of HHL (hippuryl-histidyl-leucine) to 5 ml of 0.1 M sodium borate buffer (pH 8.3) containing 0.3 M NaCl was added and reacted at 37° C. for 30 minutes. After adding 250 μl of 1 N HCl to stop the reaction, 1.5 ml of ethyl acetate was added and stirred for 15 seconds, and then centrifuged (3,000 rpm, 5 min, 4° C.) to obtain 1 ml of the supernatant. Got it. The supernatant was completely dried, 3 ml of distilled water was added and stirred, and the absorbance was measured at 228 nm with a spectrophotometer. 50 μl of distilled water was used as a negative control group, and 0.1% of captopril was used as a positive control group to compare the activity with the Examples. The ACE enzyme activity was converted into a% value of the absorbance ratio before and after addition of the sample.

ACE hydrolyzes C-terminal dipeptide (His-Leu) from angiotensin I produced by renin to produce angiotensin II, which exhibits strong vasoconstriction.

division blank captopril Example 1 Example 2 Comparative Example 1 ACE inhibitory ability (%) 100 227.4 219.2 234.8 103.4

As shown in Table 4 above, as a result of examining the ACE activity inhibition ability, it was confirmed that Examples 1 and 2 of the present invention had superior ACE activity inhibition ability compared to the control (blank) and Comparative Example 1. It was confirmed that the values of Examples 1 and 2 were similar to captopril (1 mg/ml, positive control), which is an antihypertensive agent currently on the market.

Test Example 5. AChE (Acetylcholine esterase) activity measurement

50 μl of AchE (25 mU/ml) isolated from PC12 cells was used as a reaction enzyme and 1 mM of acetylcholine iodide dissolved in 0.1 M sodium phosphate buffer (pH 8.0) was used as a substrate, and the color developing reagent was 39.6 mg of 5 ,5'-dithiobis-2-nitrobenzoic acid and 15 mg of sodium hydrogen carbonate were dissolved in 10 ml of 0.1 M sodium phosphate buffer (pH 7.0) to prepare. The enzyme reaction was carried out as follows. To 2 ml of 0.1 M sodium phosphate buffer (pH 8.0), 20 μl of a mixture of baicalin and AGW, 200 μl of a 10 mM DTNB solution, and 100 μl of enzyme (0.03 U) were added, and maintained at 37°C for 10 minutes. , 200 μl of the substrate was added and reacted for 3 minutes, and the absorbance value was measured at 412 nm with a spectrophotometer. For the activity of AChE, the ratio of absorbance before and after addition of the sample was converted into a% value. The absorbance ratio before and after was converted into a% value.

division blank Example 1 Example 2 Comparative Example 1 AChE activity (%) 100 59.8 43.6 98.7

As shown in Table 5 above, as a result of examining the activity of AChE, an enzyme that decomposes Acetylcholine to prevent smooth neurotransmission in the body, Examples 1 and 2 of the present invention were compared to the control (blank) and Comparative Example 1 AChE It was confirmed that the activity inhibiting effect was excellent.

Test Example 6. Nitric oxide (NO) production inhibition effect measurement

The amount of NO produced from LPS-induced PC12 cells was measured as the form of NO2- present in the cell culture medium using Griess reagent. PC12 cells were adjusted to 1×10 ⁵ cells/well using DMEM medium, then inoculated into a 24 well plate, and cultured in a humidified incubator at 37°C and 5% CO2. The cells were pretreated with the examples and cultured for 24 hours. The cell culture supernatant and the same amount of Griess reagent were mixed and reacted for 10 minutes in a 96 well plate, and the absorbance was measured at 540 nm and the ratio was converted to a% value.

division LPS Example 1 Example 2 Comparative Example 1 nitritE generation ability (%) 0 14.5 12.8 68.7

As shown in Table 6 above, it was confirmed that the composition of Examples 1 and 2 of the present invention inhibited NO production compared to Comparative Example 1 as to the effect of inhibiting the production of NO generated from PC12 cells by LPS (Lipopolysaccharide). .

Animal experiment

The inhibitory effect of dopaminergic cells, the main cause of Parkinson's disease, was analyzed. The amyloid beta protein was injected into brain tissue using a stereotaxic system.

5-week-old male mice (SAMTACO, Korea) were used, and 4-week-old mice were ordered and adapted for 1 week, and a stereotaxic system (KOFT, CA, USA) was used. The experiment was conducted using an adapted 5-week-old mouse. In this experiment, a model was created by injecting it into a desired area (0.0 mm posterior to bregma; 2.0 mm lateral to midline; 3.0 mm ventral to the dura) using a stereotaxic system, and specifically amyloid beta protein before proceeding with the learning experiment. Was injected through the lateral ventricle through a stereotaxic system to establish an Alzheimer's disease animal model.

Test Example 7. Passive avoidance behavior experiment

The compositions prepared in Examples and Comparative Examples were administered intraperitoneally for 2 weeks at a concentration of 5 mg/kg while the experiment was continuously conducted from 3 days after establishing an Alzheimer's disease animal model. An automated shuttle box (Model PACS-30, Columbus Instruments International Company) was used as an experimental instrument for passive avoidance behavior to confirm the protective effect of each peptide. The shuttle box is divided into two rooms of the same size (19″L x 9″W x 10.875″H) by a partition door (3″L x 2.625″W), and the floor is equipped to allow electric current to flow. Each room was illuminated by a 20W bulb placed on a hinged plexiglass lid. White paper can enter a dark room with less than 60 dB noise and dimmed lighting through the partition door. When the white paper is initially placed in a room with a light on and the partition door is opened, the room is looked around and passed to a dark room. At this time, the partition door is automatically closed and the light is turned off. This training process was continued until the white paper entered the dark room within 20 seconds. After 24 hours of completing the training process, the white paper was placed in a lighted room again, and when entering a dark room, the partition door was closed and a current (1 mA) was allowed to flow through the bottom of the shuttle box for 3 seconds. Two weeks after the lateral ventricle injection of amyloid beta protein (8 nmol/5 μl in normal saline), the white paper was placed in the illuminated room of the shuttle box and the time until it passed to the dark room was measured. The time for not entering the darkest room was set to 5 minutes. As a control, an Alzheimer's disease animal model was used.

division Normal Control Example 1 Example 2 Comparative Example 1 Passive avoidance ability
(sec) 198.1 29.4 138.5 156.9 68.5

As shown in Table 7 above, the group to which the composition for improving cognitive function prepared according to Examples 1 and 2 of the present invention was administered significantly improved the decrease in cognitive function compared to the group to which the composition of Comparative Example 1 was administered. Confirmed.

Test Example 8. Underwater maze learning memory behavior experiment

This experiment is to confirm the protective effect of each substance in the learning and memory reduction of the animal model by neurotoxic substances through the animal experiment method with aquatic rice. Before proceeding with the learning experiment, an animal model of Alzheimer's disease was established by injecting amyloid beta protein through the lateral ventricle through a stereotaxic system. Then, each substance was administered intraperitoneally for 7 days during which the underwater maze behavior experiment was continuously conducted from 3 days after surgery. First, we checked the reference test that allowed us to recognize the progress of learning and the time to recognize the surrounding environment and form memories for five days.

After making a model in which amyloid beta protein was injected, an aquatic maze experiment was started in mice injected with the compositions of Examples and Comparative Examples at a concentration of 5 mg/kg continuously for 2 weeks. A black plastic water tank was used as a cage, and a circular platform (14 x 14 cm) made of metal with a diameter of 120 cm and painted in black was placed 1 cm below the water line and the experiment was conducted. The platform divides the tank into 4 zones and is placed between the walls in the middle. Starting from each of the 4 zones of the tank divided into north, south, east, west, starting points were randomly selected. Latency time was defined as the time of arrival from the starting point to the platform. Using a computer connected to an image analyzer (SMART software basic version, Frame Grabber board, Panlab s.l., Denmark), we observed the swimming behavior in the tank. After placing the mouse in the tank for the first time, analysis and mechanical operation start 3 seconds later, and the recording is designed to stop automatically after staying on the platform for 2 seconds. In this experiment, the starting position was changed 4 times for 4 days, and the experiment was conducted under the same conditions, and the incubation time was recorded and analyzed for comparison. At this time, the analysis is analyzed as working memory, which can also be analyzed as short-term memory. And on the 5th day, under the same conditions, only the platform was removed and the number of times that the original platform existed was confirmed. It is analyzed as a reference memory and can also be analyzed as long-term memory. At this time, the standard of the longest time was 5 minutes, and the time to analyze the number of times to visit the platform was also 5 minutes. Changes in latency time indicate a decline and recovery of memory, and the longer this time, the higher the memory.

division Normal Control Example 1 Example 2 Comparative Example 1 Learning memory behavior with underwater maze
(sec) 5 day 62.3 8.4 33.2 41.8 19.3 7 day 87.5 14.2 45.9 50.3 21.9

As shown in Table 8 above, it was confirmed that the group to which the compositions prepared according to Examples 1 and 2 of the present invention were administered significantly improved cognitive decline compared to the group to which the composition of Comparative Example 1 was administered.

Moreover, as a result of checking the walking test to see how much memory is retained when the underwater maze test is conducted 24 hours after the learning has been completed for 5 days, the test speed on the 5th day and the test speed on the 7th day slightly increase, helping to maintain memory. It was confirmed that it became.

This means that the compositions of Examples 1 and 2 are effective in improving cognitive function.

Clinical examination

A total of 8 men and women over 60 o'clock with cognitive problems were recruited. Drugs or supplements that affect cognitive abilities, have a history of allergies to fish, or use drugs or supplements that cannot be tested were excluded from the study, and informed consent was obtained from all subjects.

The treatment period was 6 weeks, and all subjects took 3 capsules of the compositions of Example 1, Example 2, and Comparative Example 2 each day.

Cognitive ability was evaluated at the start of the study and at the end of the study (6 weeks) using the Clinical Dementia Assessment Scale (CDR).

Subjects participated in a training session for 1 week prior to baseline measurement and treatment initiation to conduct a sequence study. In each test session, parallel but equivalent forms of testing are provided, and the CDR contains 9 individual tasks and 5 composite elements calculated from the results of a single task.

The “attention” component is derived by combining the reaction times (simple and selective reaction times and numerical boundaries) of the three attention tasks. The “Continuity of Attention” factor is derived by calculating the combined percentage accuracy at the selected response time and numeric boundary tasks, and the “Working Memory Quality” factor is derived by combining the accuracy scores of the two working memory tests (spatial and numeric working memory). , The “Quality of One-Time Secondary Memory” factor is derived by combining the composite score working memory with the percent accuracy score of all secondary memory tests (recognition and recall tests). The "memory speed" factor is derived by combining the response times of the four memory operations (delayed picture and word recognition, numeric task memory and spatial memory).

In the statistical method, P, 0.05 was considered significant without correction for multiple comparisons. Analysis was performed using SPSS software (version 17; SPSS Inc, Chicago, IL).

division Start of research Example 1 Example 2 Comparative Example 1 Immediate word recall (%) 36.15±3.81 39.17±2.69 40.96±3.54 37.44±4.15 Simple reaction time (ms) 347.56±21.57 330.67±20.15 321.07±17.51 341.61±19.68 Numeric Awakening Target Detection (%) 94.31±0.84 96.56±0.78 97.11±0.81 95.15±0.70 Numeric Awakening False Alarm (#) 2.00±0.49 1.54±0.53 1.47±0.47 1.89±0.31 Numeric Awakening Response Time (ms) 466.28±16.48 483.56±15.16 492.68±11.25 471.45±14.84 Selection reaction time accuracy (%) 93.55±1.12 95.18±1.10 96.44±0.72 94.28±0.84 Selective reaction time (ms) 557.26±17.46 540.15±10.59 531.67±11.42 551.62±12.85 Spatial working memory sensitivity index (#) 0.97±0.01 0.97±0.02 0.96±0.01 0.96±0.01 Space working memory response time (ms) 1202.05±98.45 1126.45±64.38 1085.85±74.31 1181.27±56.96 Delayed word recall (%) 20.00±3.84 27.85±3.10 30.09±2.71 23.56±2.94 Word recognition sensitivity index (#) 0.61±0.05 0.74±0.03 0.80±0.04 0.69±0.04 Word recognition response time (ms) 1048.17±68.48 1197.62±75.42 1288.29±79.10 1101.54±81.41 Photo recognition sensitivity index (#) 0.74±0.04 0.85±0.03 0.91±0.04 0.79±0.02 Photo recognition response time (ms) 1201.00±68.72 1269.46±51.08 1307.25±65.75 1237.52±72.11

As shown in Table 9 above, the compositions prepared according to Examples 1 and 2 of the present invention were found to improve cognitive ability compared to Comparative Example 1.

Hereinafter, examples of the formulation of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

500 mg of the composition obtained in Example 1

100 mg lactose

10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablet

300 mg of the composition obtained in Example 1

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet preparation method.

Formulation Example 3. Preparation of Capsule

200 mg of the composition obtained in Example 1

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

600 mg of the composition obtained in Example 1

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared with the above ingredients per ampoule according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

4 g of the composition obtained in Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid formulation, add and dissolve each component in purified water, add an appropriate amount of lemon flavor, mix the above ingredients, add purified water, add purified water and adjust the total to 100 g, then fill in a brown bottle for sterilization. To prepare a solution.

Formulation Example 6. Preparation of granules

1,000 mg of the composition obtained in Example 1

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

1.7 mg of nicotinic acid amide

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

1.75 mg ferrous sulfate

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules, but it is possible to arbitrarily modify the mixing ratio. After mixing the above ingredients according to a conventional granule preparation method, granules And can be used to prepare a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of functional beverage

1,000 mg of the composition obtained in Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 mL with purified water

After mixing the above ingredients according to a normal health drink manufacturing method, stirring and heating the resulting solution at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and stored in a refrigerator. It is used to prepare the functional beverage composition of the invention.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the purpose of use.

Claims (9)

A phospholipid mixture comprising phosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidic acid;
Neutral lipid mixtures including EPA and DHA;
Vitamin mixtures including vitamin E, vitamin C, vitamin D and vitamin B12; And
Including;
The phospholipid mixture comprises 10 to 50 parts by weight of phosphatidylcholine, 20 to 50 parts by weight of phosphatidylethanolamine, 5 to 30 parts by weight of phosphatidylinositol, and 5 to 30 parts by weight of phosphatidylinositol and phosphatidylcholine. (phosphatidic acid) contains 20 to 50 parts by weight,
It is contained in an amount of 0.1 to 10 parts by weight of a neutral lipid mixture, 0.1 to 5 parts by weight of a vitamin mixture, and 1 to 10 parts by weight of an oil and fat based on 100 parts by weight of the phospholipid mixture,
The fat and oil are fish oil consisting of medium chain triglyceride (MCT) oil; Perilla oil, evening primrose oil, blackcurrant oil, borage oil, hemp seed oil, cottonseed oil, safflower oil, soybean oil, and one or more vegetable oils selected from the group consisting of sunflower oil; Cognitive function improvement composition, characterized in that it is a mixed oil. delete delete The composition for improving cognitive function according to claim 1, wherein the neutral lipid mixture contains 25 to 100 parts by weight of DHA based on 100 parts by weight of EPA. The composition for improving cognitive function according to claim 1, wherein the vitamin mixture is a mixture of vitamin E, vitamin C, vitamin D, and vitamin B12 in a weight ratio of 1: 0.5-2: 0.1-1: 1-10. According to claim 1, wherein the vitamin E is D-alpha-tocopherol (D-α-Tocopherol) based on 100 parts by weight of DL-alpha-tocopherol (DL-α-Tocopherol) 10 to 80 parts by weight, D-alpha-tocopherol Peryl Acid Succinate (D-α-Tocopheryl Acid Succinate) 5 to 70 parts by weight, D-alpha-Tocopheryl Acetate (D-α-Tocopheryl Acetate) 10 to 70 parts by weight and DL-alpha-Tocopheryl Acetate (DL-α -Tocopheryl Acetate) Cognitive function improving composition, characterized in that the mixture containing 5 to 70 parts by weight. delete A food composition for improving brain function comprising the composition of any one of claims 1 and 4 to 6. A pharmaceutical composition for the prevention and improvement of Alzheimer's, comprising the cognitive function composition of any one of claims 1, 4 to 6 as an active ingredient.