KR102107304B1 - Natural liposome comprising red yeast rice, process for the preparation thereof, and food composition comprising the same - Google Patents

Abstract

The present invention is a liposome-constituent material containing one or more natural emulsifiers, natural-derived solvents, natural coatings, and natural preservatives, and further comprises liposomes comprising one or more selected from the group consisting of sphingolipids, sterols, and glycoproteins. matter; And a gaba, gongmyeong-tang, or a mixture thereof as a collection material, and relates to a natural liposome formed by mixing the natural emulsifier, the natural-derived solvent, and the gaba, gongmyeong-tang, or a mixture thereof.
In addition, the present invention relates to a method for preparing the natural liposome and a food composition comprising the same.

Description

Natural liposomes, manufacturing method thereof, and food composition comprising the same {NATURAL LIPOSOME COMPRISING RED YEAST RICE, PROCESS FOR THE PREPARATION THEREOF, AND FOOD COMPOSITION COMPRISING THE SAME}

The present invention relates to a natural liposome, a method of manufacturing the same, and a food composition comprising the same.

Gaba (red yeast rice: gamma-aminobutyric acid) is fermented after inoculation of gaba bacteria ( Monascus sp.) By steaming rice, and contains several beneficial metabolites produced during fermentation. Among the ingredients in Gaba, monacolin K is known to inhibit the biosynthesis of cholesterol in the liver, and to increase the level of high density (HDL) cholesterol while lowering the level of low density (LDL) cholesterol. In addition, Gaba contains gamma aminobutyric acid, which suppresses the accumulation of cholesterol and triglycerides in blood vessels, thereby helping blood flow smoothly and reducing blood pressure.

As such, foods, health supplements, and medicines using gaba are actively being developed to utilize various effects of gaba. Korean Patent Publication No. 2014-0037294 discloses a persimmon chip fermented by inoculation of gaba bacteria on persimmon, and Korean Patent Publication No. 2013-0029890 discloses a composition containing monascin extracted from gaba bacteria and its preparation Methods are disclosed.

Also, Chongmyeongtang is known as a herbal material related to brain function improvement. Chongmyeongtang is a herbal medicine ingredient mainly composed of Boksin, Wonji and Seokchangpo.

However, when ingesting a product containing gaba or gongmyeong-tang, the effect of gaba or gongmyeong-tang can be expected, but the absorption rate in the body is different for each individual, so the effect may be different. Accordingly, there is a need to develop a technique for increasing the absorption of gaba and gongmyeongtang in the body so as to exert an excellent effect when ingesting a gaba-containing product or gongmyeongtang-containing product.

Korean Patent Publication No. 2014-0037294 Korean Patent Publication No. 2013-0029890

An object of the present invention is to provide a natural liposome that can be easily injected into a living body while being harmless to a human body and exhibits an excellent memory enhancement and learning improvement effect, a manufacturing method thereof, and a food composition comprising the same.

An example of the present invention is a liposome-constituent material containing at least one natural emulsifier, natural-derived solvent, natural coating agent, and natural preservative, and further contains at least one selected from the group consisting of sphingolipids, sterols, and glycoproteins. Liposome-constituent substance; And it includes a gaba, gongmyeongtang, or a mixture thereof as a collection material, and provides a natural liposome formed by mixing the natural emulsifier, the natural-derived solvent, and the gaba, gongmyeongtang, or a mixture thereof by ultrasonic treatment.

Another example of the present invention is (a) a first step of mixing and sonicating a natural-derived solvent with one or more natural emulsifiers at 35 to 80 ° C; (b) a second step of mixing and ultrasonicating one or more selected from the group consisting of sphingolipids, sterols, and glycoproteins in the mixture of the first step; (c) a third step of adding gaba, gongmyeongtang, or a mixture thereof to the mixture of the second step and ultrasonically mixing the mixture; (d) a fourth step of mixing and sonicating the mixture of the third step with a natural coating at 35 to 80 ° C; And (e) a fifth step of adding a naturally occurring solvent and a natural preservative to the mixture of the fourth step and subjecting it to ultrasonic treatment.

Another example of the present invention provides a food composition comprising the natural liposome.

The natural liposomes according to the present invention can provide an effect of improving memory by Gaba, Chong Myung-Tang, forgetfulness, brain metabolism promotion, nerve suppression, fatigue recovery, etc.

1 is a graph comparing the absorption rate of Example 1 and Comparative Example 1, the control.
2A and 2B are graphs comparing the absorption rates of Example 2 and Comparative Example 2, and the control, respectively.
3A and 3B are graphs comparing the absorption rates of Example 3 and the control, respectively.
4A and 4B are graphs comparing the absorption rates of Example 4 and the control, respectively.
5 is a graph of absorbance measurement results by concentration according to Experimental Example 3-1.
6 is a graph showing the results of measuring the cell protection effect of each sample.
7 and 8 show the results of confirming the Gaba receptor and serotonin receptor activity of neurons by the western blot method according to each sample.
9 is a graph showing the results of measuring the cell protection effect of each sample.

Hereinafter, a natural liposome according to the present invention, a method for manufacturing the same, and a food composition comprising the same will be described in more detail with reference to the accompanying drawings. However, these descriptions have been presented by way of example only to help understanding of the present invention, and the scope of the present invention is not limited by these exemplary descriptions.

<Natural liposome>

The liposome according to an embodiment of the present invention is a liposome-constituent material containing one or more natural emulsifiers, natural-derived solvents, natural coatings, and natural preservatives, and one or more selected from the group consisting of sphingolipids, sterols, and glycoproteins. Liposome-constituent substances further containing; And it includes a gaba, gongmyeongtang, or a mixture thereof as a collection material, and provides a natural liposome formed by mixing the natural emulsifier, the natural-derived solvent, and the gaba, gongmyeongtang, or a mixture thereof by ultrasonic treatment. The natural liposomes are distinguished from conventional synthetic liposomes in that they do not contain synthetic ingredients.

In the present invention, 'natural liposomes' refers to liposomes in which all components except the trapping material are made of natural components only. In the case of the trapping material contained in the natural liposome, it may be composed of synthetic components, but in the case of the trapping material, it is preferably made of natural components.

1. Liposomal components

The liposomes of the present invention are natural liposomes that include natural emulsifiers, naturally derived solvents, natural coatings and natural preservatives. Natural liposomes made of these natural substances are harmless because they are non-toxic to the human body and can be ingested. In addition, the liposomes can form a stable phospholipid bilayer, thereby effectively delivering trapping substances to cells.

[Natural emulsifier]

Natural emulsifiers are emulsifiers derived from natural substances and constitute a lipid bilayer. The natural emulsifier includes phospholipids and fatty acids. Phospholipids included in natural emulsifiers are phospholipids derived from nature, and fatty acids are also natural fatty acids derived from nature.

Phospholipids are contained in egg yolk, soybean, etc., and can be obtained by extraction and separation and purification from them. The phospholipids include phosphatidyl choline (lecithin), lysophosphatidyl choline, phosphatidyl ethanolamine, phosphatidic acid, phosphatidyl serine, phosphatidyl glycerol (phosphatidyl glycerol) , Phosphatidyl inositol and one or more natural phospholipids selected from the group consisting of hydrogenated products (for example, hydrogenated phosphatidyl choline-derived from soybean) may be used. Postatidyl choline, lysophosphatidyl choline or hydrogenated phosphatidyl choline with good stability may be used.

The fatty acids are contained in a lot of nuts such as walnuts, pecans, almonds, pine nuts, seeds such as olives, corn, flax, cotton, rapeseed, evening primrose, and can be obtained by extraction and separation and purification. As the fatty acid, at least one fatty acid selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid may be used. .

The natural emulsifier may include 70 to 90% by weight of natural phospholipid and 10 to 30% by weight of natural fatty acid based on 100% by weight of the total content of natural phospholipid and natural fatty acid. The natural emulsifier may include the remaining amount of other solvents. As a solvent, it is preferable to use a solvent derived from nature, and it is more preferable to use distilled water.

The natural emulsifier can be used by mixing two or more. For example, when two types of natural emulsifiers are used in combination, for example, as the first natural emulsifier, phosphatidyl choline, lysophosphatidyl choline, phosphatidyl ethanolamine, and palmitic acid (palmitic acid), stearic acid, oleic acid, linoleic acid, linolenic acid (derived from soybeans) may be used, and the second natural emulsifier is hydrogenated phosphatidyl choline (hydrogenated) phosphatidyl choline) can be used. In addition, when three kinds of natural emulsifiers are mixed and used, for example, in addition to the first natural emulsifier and the second natural emulsifier, cetearyl olivate and sorbitan oleate as third natural emulsifiers ( sorbitan olivate) can be used.

In addition, the natural emulsifier may further include natural fatty acids. Evening primrose oil may be used as the natural fatty acid. The content of the natural fatty acid may be 0.1 to 5% by weight based on the weight of the total natural emulsifier.

The content of the natural emulsifier is preferably 0.1 to 10% by weight based on the weight of all natural liposomes.

[Natural derived solvent]

The naturally-derived solvent causes the lipid bilayer of the natural emulsifier to form a capsule surrounding the capture material.

As the solvent, one or more selected from the group consisting of a naturally-derived solvent, for example, distilled water, naturally-derived glycerin, natural ethanol, natural propanediol, natural glycerin, and fermentation alcohol may be used. Preferably, distilled water, naturally occurring glycerin, and natural ethanol can be used, and more preferably distilled water can be used.

The content of these solvents is preferably a residual amount excluding natural emulsifier, natural coating agent, natural preservative, trapping material, sphingolipid, sterol, and glycoprotein based on the weight of the total natural liposome.

[Natural preservatives]

Natural preservatives are substances that maintain the stability of the final natural liposomes by preventing decay of the liposome-constituent substances (natural emulsifiers, naturally-derived solvents, natural coatings) and trapping substances present in the liposomes.

The natural preservative may be used without limitation as long as it is a natural preservative that can be added to food or pharmaceuticals. For example, natural extracts can be used. One or more selected from the group consisting of citrus extract, citron extract, eucalyptus extract, ginseng extract, clove extract and grapefruit extract may be used. Preferably, citrus extract and grapefruit extract may be used. The preservative is dehydroacetic acid, sorbic acid, benzoic acid, methyl paraoxybenzoate, and propanoic acid, which are commonly used as food preservatives in that they are natural preservatives. ) And their salts and chemical preservatives.

The content of the natural preservative is preferably 0.001 to 3% by weight, more preferably 0.01 to 0.3% by weight, based on the total weight of the natural liposome.

[Other liposome-constituent substances]

In addition to the above components, as a liposome-constituting material surrounding the trapping material, one or more substances selected from the group consisting of spingolipid, sterols, and glycoproteins are included.

Unlike conventional natural liposomes, which have a structure in which the trapping material is wrapped with a natural emulsifier that is a substance similar to the skin, that is, a phospholipid, in one example of the present invention, in addition to the natural emulsifier, sphingolipid, sterol, which is a substance similar to a biofilm component, At least one material selected from the group consisting of glycoproteins has a structure surrounding the additional trapping material. Therefore, the trapped material can be better absorbed by the mucous membrane of the small intestine.

The sphingolipids include, for example, sphingomyelin, and the sterols include, for example, phytosterols that are vegetable sterols, campus terrols, and vega pure.

Sphingomyelin is a type of sphingolipid and has a form in which phosphocholine is attached to the -OH portion of sphingosine. Sphingomyelin has a structure similar to phosphatidylcholine, so it can form a double layer with phosphatidylcholine. Sphingomyelin is a component of the cell membrane, which is important for the function of maintaining the plasma membrane and also has the ability to regulate various cell metabolism. As sphingomyelin, milk ceramide is mentioned, for example.

There is no particular limitation on the content of the sphingolipids used, but when considering the absorption rate, 0.5 to 5% by weight is preferable based on the total weight of the natural liposome, and 1 to 3% by weight is more preferable.

Sterol (sterol) refers to an alcohol having a steroid skeleton that is widely distributed in animal and plant systems. It is also called sterin. Representative animal sterols include cholesterol and cosprostanol, phytosterols and vega pure as plant sterols, and ergosterols as fungal sterols. Other major natural sterols include chimosterol, epicoprostanol, 24-dehydrocholesterol, 7-dehydrocholesterol, cerebrosterol, ascosterol, pecosterol, and campus terrol. Sterols are basically four rings and are isoprenoid-based hydrocarbons. Cholesterol is amphiphilic due to the large number of -OH groups, and does not belong to phospholipids because it has no phosphate structure. Because cholesterol and other sterols are very hydrophobic, it is impossible to form a cell membrane with them alone. Instead, it enters between the phospholipid membranes and prevents the phospholipids from sticking too much, thereby maintaining its fluidity and at the same time enhancing physical rigidity. Do it. Cholesterol is also used in cell membranes, but is also used to emulsify fat (in gallbladder) when digesting fat, steroid hormones, and vitamin D synthesized in the skin or kidneys. In addition, cholesterol is used as a signal transduction molecule that plays an important role in embryonic differentiation by covalently binding to the Hedgehog protein.

The content of sterol used is not particularly limited, but when considering the absorption rate, 0.01 to 2% by weight is preferred based on the total weight of natural liposomes, 0.05 to 1.5% by weight is more preferable, and 0.08 to 0.8% by weight is most preferred. .

Glycoproteins play a role in helping the intestinal mucosa to permeate, for example, carrot collagen. By incorporating glycoproteins, the intestinal mucosal permeability can be increased.

There is no particular limitation on the content of the glycoprotein used, but when considering the absorption rate, 2 wt% or more is preferred based on the total weight of the natural liposome, more preferably 2.5 wt% or more, and most preferably 5 wt% or more.

The ratio of emulsifier, sphingolipid, sterol, and glycoprotein can be appropriately adjusted within the scope of the present invention. Preferably, the sphingolipids, sterols, and glycoproteins based on the total weight of natural liposomes include 1.0 to 10% by weight in total.

In addition, when all three components are included, preferably, the ratio of sphingolipids: sterols: glycoproteins is 2: 0.1 to 0.5: 1 to 5 based on weight, and more preferably 2: 0.2 to 0.4: 2 to 4.

2. Collection material

The capture material according to the present invention is a material that is collected inside the lipid bilayer of the liposome containing the natural emulsifier, natural solvent, natural coating agent and natural preservative. Gaba may be used as the collection material.

Gaba contains beneficial metabolites produced during the fermentation process by Gaba, and in particular, it has been known that the effect of improving blood lipid composition and blood circulation by Monacholine K is known. These gaba are processed in powder or liquid form to facilitate ingestion, and are used as a collection material of liposomes to increase the absorption rate in the body. Preferably, Gaba powder can be used as a collection material. At this time, in order to increase the absorption rate in the body while preventing the functional components of the gaba extract from being destroyed, the molecular size may be reduced by ultrasonic treatment. These gaba extracts may be sufficiently dissolved and used in butylene glycol, propanediol, glycerin, fermented alcohol, etc. derived from natural substances so that they can be easily mixed with liposome-constituent substances.

Gaba helps lower blood pressure, improves brain metabolism and prevents dementia and helps improve memory. Gaba has a mental stabilizing effect, which helps insomnia and is effective in autonomic nerve disorders. It promotes liver function activity and alcohol metabolism, so it is effective in removing hangovers and improves kidney and liver functions and prevents obesity. As such, Gaba has a neuro-suppressive action and a psycho-stable function, and has an effect of promoting metabolism of the brain, and thus is receiving attention as a functional food material.

Chongmyeongtang can also be used as a collection material. Chongmyeongtang is known as a herbal material related to brain function improvement. Chongmyeongtang is a herbal medicine ingredient mainly composed of Boksin, Wonji and Seokchangpo.

Chongmyeongtang is obtained by dipping Baekboksin, the original paper in licorice water, removing the planted plant, making it into persimmon juice, and then slicing changchangpo. Alternatively, water is added to raw paper, baekboksin, cheonma, licorice, jujube, and whole raw steel as ingredients, and extracted at a first stage temperature, for example, 75 to 85 degrees Celsius for a certain period of time, for example, 2 to 4 hours. , Two-step temperature, for example, from 85 to 95 degrees Celsius for a certain period of time, for example, 4 to 6 hours is produced by a method such as extraction and filtering.

Chongmyeong-tang is used to treat conditions such as memory loss and forgetfulness. Chongmyeong-tang helps promote brain cell metabolism, restores memory, and promotes long-term memory activity by inhibiting the activity of enzymes that break down neurotransmitters. It suppresses stress accumulation and is effective in recovering fatigue. In addition, Chongmyeongtang is effective in increasing brain blood flow and protecting brain neurons.

The content of the gaba extract, cheonmyeongtang, or a mixture thereof is preferably 0.1 to 30% by weight based on the total weight of the natural liposome.

3. Other

As a component for coating other capture materials, a coating agent may be additionally included. As the coating agent, a natural coating agent is preferably used. Dietary fiber, starch, polysaccharide, etc. may be used as the coating agent, for example, maltodextrin, tapioca, guar gum, corn starch, etc. may be used. It is preferable to use maltodextrin from the viewpoint of high stability so that the trapped material is well absorbed.

As described above, the natural liposome of the present invention is a capsule form surrounded by a lipid bilayer surrounding a gaba extract or gongmyeongtang having a small particle size, and the gaba extract or gongmyeongtang can be easily delivered into cells. When the natural liposomes are delivered to the cells, the lipid bilayer of the liposomes and the cell membrane are easily combined and the cell uptake rate of the gaba extract or myeongmyeong-tang increases, thereby improving memory by gaba or myeongmyeong-tang, improving forgetfulness, promoting brain metabolism, and suppressing nerves. , Can provide effects such as fatigue recovery.

<Production method of natural liposomes>

The present invention provides a method for preparing natural liposomes by treating ultrasonic waves so that the capture material and the liposome constituent materials are uniformly mixed. However, it is not limited only by this manufacturing method, and the steps of each process may be modified or selectively mixed as necessary.

A preferred method for preparing a natural liposome according to an embodiment of the present invention includes: (a) a first step of mixing a natural-derived solvent with one or more natural emulsifiers at 35 to 80 ° C. and treating with ultrasound; (b) a second step of mixing and sonicating sphingomyelin, sterol, and glycoproteins in the mixture of the first step; (c) a third step of adding gaba, gongmyeongtang, or a mixture thereof to the mixture of the second step and ultrasonically mixing the mixture; (d) a fourth step of mixing and sonicating the mixture of the third step with a natural coating at 35 to 80 ° C; And (e) a fifth step of adding a naturally-derived solvent and a natural preservative to the mixture of the fourth step and ultrasonicating the mixture. After the fifth step, the sixth step of repeatedly ultrasonic treatment while gradually lowering the temperature may be further included.

Hereinafter, the manufacturing method will be described by dividing each step by step as follows.

(a) Step 1

This step is a step of mixing and sonicating a natural-derived solvent with one or more natural emulsifiers at 35 to 80 ° C.

Distilled water may be used as the natural-derived solvent, or natural ethanol, natural propanediol, natural glycerin, fermented alcohol, etc. derived from natural substances may be used.

As the natural emulsifier, one or more natural emulsifiers containing phospholipids and fatty acids derived from natural substances may be used.

The ultrasonic treatment is an operation for homogenizing a mixture of a natural-derived solvent and a natural emulsifier, and may use an ultrasonic liquid processor.

These natural-derived solvents and natural emulsifiers are heated to 35 to 80 ° C., preferably 50 to 75 ° C., more preferably 60 to 70 ° C., and ultrasonically treated to homogenize the phospholipids and fatty acids of the natural emulsifier in the naturally-derived solvent. You can. The temperature range is an advantageous temperature range for the emulsifier to stably form liposomes.

(b) Stage 2

This step is a step of mixing and ultrasonicating one or more types selected from the group consisting of sphingolipids, sterols, and glycoproteins in the mixture of the first step.

First, the sphingolipids are dissolved in distilled water, and then added to the mixture of the first step. Dispersed and homogenized using an ultrasonic digester.

Then, the sterol is dissolved in distilled water, and then added to the first stage mixture in which the sphingolipid is dissolved. Dispersed and homogenized using an ultrasonic digester.

Then, the glycoprotein is dissolved in distilled water, and then added to the first stage mixture in which sterol and sphingolipid are dissolved. Dispersed and homogenized using ultrasonic waves.

(c) Stage 3

This step is a step of adding gaba, gongmyeongtang, or a mixture thereof to the mixture of the second step and mixing by ultrasonic treatment.

The gaba extract or gongmyeongtang can be obtained by crushing the gaba or gongmyeongtang or extracting it in a liquid state at a temperature, pressure, and humidity condition where the functional components are not destroyed. At this time, the method of grinding the gaba can be used without limitation, conventional grinding methods known in the art. In addition, as a method of extracting gaba and cheonmyeongtang in a liquid state, a conventional extraction method known in the art can be used without limitation. As an example, extracts in liquid or powder form may be obtained using extraction methods such as cold needle extraction, ultrasonic extraction, reflux cooling extraction, and hot water extraction. The gaba extract and Chongmyeong-tang can be used in a form dissolved in butylene glycol, propanediol, glycerin, fermented alcohol, etc. derived from natural substances to be uniformly mixed with the mixture of the first step. When the gaba extract and gongmyeong-tang are dissolved in fermented spirits, the mixing ratio of the gaba extract and fermented spirits is gaba extract (or gyeongmyeong-tang) based on weight ratio: 0.1 to 1: 1, and preferably 0.3: 1.

Through the third step, each component can be mixed homogeneously and liposomes having a small particle size can be formed. The sonication can be performed, for example, after adding the gaba extract to the mixture of the second step and heating to 35 to 80 ° C. to mix. However, when the mixture of the second step maintains the temperature range, ultrasonic treatment may be performed without a separate heating process.

(d) Step 4

This step is a step of mixing and sonicating the mixture of the third step with a natural coating at 35 to 80 ° C. The sonication can be performed, for example, after adding a natural coating to the mixture of the third step and heating to 35 to 80 ° C. to mix. However, when the mixture of the third step maintains the temperature range, ultrasonic treatment may be performed without a separate heating process.

The natural coating agent helps the body absorb the natural liposomes that are finally produced.

(e) Step 5

This step is a step of adding a naturally-derived solvent and a natural preservative to the mixture of the fourth step and sonication. It is also possible to preheat the naturally-derived solvent and the natural preservative to a specific temperature, and then perform ultrasound.

The same solvent used in the first step may be used as the naturally-derived solvent.

The natural preservative maintains the stability of the natural liposomes by preventing the decay of the liposome-constituent substances (natural emulsifiers, natural-derived solvents, natural coatings) and the trapping substances present in the liposomes. In addition, the natural preservative is derived from natural materials and is non-toxic, harmless to the human body, and does not damage natural liposomes.

(b) second step, (c) third step, (d) fourth step. (e) The ultrasonic treatment in the fifth step is performed at 35 to 80 ° C, preferably at 60 to 70 ° C.

(f) Step 6

This step is a step of repeatedly ultrasonicating while gradually lowering the temperature after the fifth step. This step can optionally be included.

The natural liposomes prepared by the above preparation method gradually lower the temperature, preferably by finally ultrasonically treating at 35 to 70 ° C. for 1 to 10 minutes, preferably 3 to 8 minutes, to thereby form the particles formed in the fifth step. Smaller, more uniform final natural liposomes are prepared.

The ultrasonic treatment may be repeatedly performed to prepare natural liposomes having a desired particle size, and may preferably be performed 1 to 3 times.

As described above, the method for producing a natural liposome of the present invention forms a liposome with a natural material, which is harmless to the human body, and forms a natural liposome containing a small particle size gaba extract, total myeongtang to increase the absorption rate of gaba, myungtang for cells. You can.

<Food composition containing natural liposomes>

An example of the present invention provides a food composition comprising the natural liposome as an active ingredient.

Natural liposomes according to an embodiment of the present invention by combining with the cell membrane to increase the cell uptake rate of gaba extract or gongmyeongtang, so that the components of gaba, gongmyeongtang can effectively work in the human body. When the natural liposomes are ingested, the effect of gaba absorbed in the intestine and gongmyeongtang may appear, and thus can be used as a food composition containing natural liposomes as an active ingredient.

These food compositions are based on total weight. 0.1 to 10% by weight of the natural emulsifier, 0.001 to 3% by weight of the natural coating agent, 0.001 to 3% by weight of the natural preservative, 0.1 to 30% by weight of the gaba, myeongtang, or mixtures thereof, the sphingolipids, sterols, sugar It may include a total of 1.0 to 10% by weight of the protein and the residual amount of the solvent derived from the nature.

The food composition of the present invention can be used without limitation on the type of food, and can be used as an additive to general foods or as a dietary supplement.

Food compositions containing such natural liposomes as active ingredients help improve memory and improve learning.

Hereinafter, the present invention will be specifically described through examples, but the following examples are only illustrative of one form of the present invention, and the scope of the present invention is not limited by the following examples.

1. Test of absorption of liposomes using synthetic gaba

[Example 1] Preparation of natural liposomes containing 99% of gaba

Natural liposomes were prepared according to the composition of Table 1 below.

division order Raw material name Content (% by weight) A One Distilled water 30.0 2 DS-Soya PC60 IP 2.5 B 3 Milk ceramide 2.0 4 Distilled water 2.5 C 5 Phytosterol 0.3 6 Distilled water 2.5 D 7 Carrot Collagen 3.0 8 Distilled water 2.5 E 9 Synthetic GABA
(99% GABA-containing powder) 5.0 10 Distilled water 2.5 F 11 Maltodextrin 1.0 G 12 Distilled water 46.15 H 13 Grapefruit extract 0.05

Step A: First, distilled water was weighed and heated to 65 ° C, and then DS-Soya PC60 IP was added to the distilled water as a natural emulsifier. It was disassembled for 10 seconds using a sonicator and stopped for 3 seconds to homogenize for a total of 30 seconds.

Step B: The milk ceramide was completely dissolved in distilled water, and then slowly added to the result of Step A completely dissolved. It was disassembled for 10 seconds using a sonicator, stopped for 3 seconds, and homogenized by dispersing for a total of 30 seconds.

Step C: The phytosterol was completely dissolved in distilled water, and then slowly added to the result of Step B completely dissolved. It was disassembled for 10 seconds using a sonicator, stopped for 3 seconds, and homogenized by dispersing for a total of 30 seconds.

Step D: Carrot collagen was completely dissolved in distilled water, and then slowly added to the result of step C completely dissolved. It was disassembled for 10 seconds using a sonicator, stopped for 3 seconds, and homogenized by dispersing for a total of 30 seconds.

Step E: GABA sigma 99% powder was completely dissolved in distilled water, and then slowly added to the result of step D completely dissolved. It was disassembled for 10 seconds using a sonicator, stopped for 3 seconds, and homogenized by dispersing for a total of 30 seconds.

Step F: The maltodextrin was added to the result of the completely dissolved step E, mixed, and heated to 65 ° C. It was disassembled for 10 seconds using a sonicator, stopped for 3 seconds, and homogenized by dispersing for a total of 30 seconds.

Step G: Distilled water was added to the result of the step F completely dissolved. It was disassembled for 10 seconds using a sonicator, stopped for 3 seconds, and homogenized by dispersing for a total of 30 seconds.

The process of dissociating for 10 seconds using a sonicator, stopping for 3 seconds, and dispersing for a total of 30 seconds was repeated once more.

Step H: Grapefruit seed extract was added to the result of step G completely dissolved.

[Comparative Example 1] Preparation of natural liposomes containing 99% of gaba

Natural liposomes were prepared according to the composition in Table 2 below in the same manner as in Example 1, except that steps B, C, and D were omitted and distilled water was used at 58.95% by weight in the last step.

division order Raw material name Content (% by weight) A One Distilled water 30.0 2 DS-Soya PC60 IP 2.5 E 3 Synthetic GABA
(99% GABA powder) 5.0 4 Distilled water 2.5 F 5 Maltodextrin 1.0 G 6 Distilled water 58.95 H 7 Grapefruit extract 0.05

[Experimental Example 1] Characteristic experiment-skin absorption rate experiment

Skin absorption was confirmed using a Franz diffusion cell (Permegear, V3A-02, USA) using a pig small intestine mucosa.

First, pig skin tissue was fixed between the donor and receptor phases. Samples of Example 1, Comparative Example 1 and control were administered to the donor of the prepared Franz diffusion cell, and the temperature was maintained at about 36.5 ° C using a constant temperature water bath. Absorbance was measured by collecting 0.25 ml of the receptor phase at 6 hour intervals. The results are shown in FIG. 1.

[Experiment result]

As shown in Figure 1, the natural liposome of Example 1 was found to have a higher absorption rate than the control of Gaba powder, a normal natural liposome (Comparative Example 1).

2. Characterization of liposomes using natural gaba

[Example 2] Preparation of natural liposomes containing 90% of gaba

GABA sigma 99% powder 5% by weight, instead of using 46.15% by weight of distilled water in step G, natural-derived GABA (fermented barley extract GABA 90% powder 5.7% by weight, in step G, 45.45% by weight of distilled water was used. Except for the same content as Table 1, natural liposomes were prepared in the same manner as in Example 1.

[Example 3] Preparation of natural liposomes containing 20% Chongmyeongtang

GABA sigma 99% powder 5% by weight, and instead of using 46.15% by weight of distilled water in step G, 20% by weight of Myeongmyeong-tang and 31.15% by weight of distilled water in step G were used in the same amount as in Table 1. Natural liposomes were prepared in the same manner as in Example 1.

[Example 4] Preparation of natural liposomes containing 10% Chongmyeongtang

GABA sigma 99% powder 5 wt%, instead of using 46.15 wt% of distilled water in step G, 10% by weight, respectively, and distilled water in step G were used in the same amount as in Table 1, except that 41.15 wt% was used. Natural liposomes were prepared in the same manner as in Example 1.

 [Comparative Example 2] Preparation of natural liposomes containing 90% of gaba

Natural liposomes were prepared according to the composition in Table 3 below in the same manner as in Example 2, except that steps B, C, and D were omitted, and 58.95% by weight of distilled water was used in the last step.

division order Raw material name Content (% by weight) A One Distilled water 30.0 2 DS-Soya PC60 IP 2.5 E 3 Naturally derived GABA
(Fermented barley extract powder, GABA 90%) 5.7 4 Distilled water 2.5 F 5 Maltodextrin 1.0 G 6 Distilled water 58.95 H 7 Grapefruit extract 0.05

[Comparative Example 3] Preparation of natural liposomes containing 20% Chongmyeongtang

Natural liposomes were prepared according to the composition in Table 4 below in the same manner as in Example 4, except that steps B, C, and D were omitted, and 43.95% by weight of distilled water was used in the last step.

division order Raw material name Content (% by weight) A One Distilled water 30.0 2 DS-Soya PC60 IP 2.5 E 3 Myeongmyeongtang 20.0 4 Distilled water 2.5 F 5 Maltodextrin 1.0 G 6 Distilled water 43.95 H 7 Grapefruit extract 0.05

 [Experimental Example 2] Characterization

(1) particle size analysis

The particle shape and size of the natural liposomes of GABA, Chongmyeong-tang, Examples 2, 3, and Comparative Examples 2 and 3 were analyzed using a scanning electron microscope (Scanning Electron Microscope S-1700). The sample was lyophilized at 60 degrees Celsius and then thinly spread and coated once with a PdPt mixed catalyst to measure.

The particle size of the sample was measured using a laser particle size analyzer (Electrophoretic Light Scattering Spectrophotometer ELS-8000, Otsuka) and the particle size distribution was analyzed.

The results are shown in Tables 5 and 6 below.

Item Particle size (nm) GABA 183.7 Comparative Example 2 71.3 Example 2 137.4

Item Particle size (nm) Myeongmyeongtang 374.8 Comparative Example 3 66.5 Example 3 213.1

As shown in Tables 5 and 6, the natural liposomes according to an example of the present invention (Examples 2 and 3) have a larger particle size compared to the normal natural liposomes (Comparative Examples 2 and 3), but compared to Gaba powder and Baekmyeongtang. Showed a small particle size.

(2) Skin absorption rate experiment

Skin absorption was confirmed using a Franz diffusion cell (Permegear, V3A-02, USA) using a pig small intestine mucosa.

First, pig skin tissue was fixed between the donor and receptor phases. Samples of Examples 2 to 4 and Comparative Example 2 and control were administered to the donor of the prepared Franz diffusion cell, and the temperature was maintained at about 36.5 ° C using a constant temperature water bath. Absorbance was measured by collecting 0.25 ml of the receptor phase at 6 hour intervals. The results are shown in FIGS. 2A, 3A, and 4A.

[Experiment result]

As shown in Figure 2a, the natural liposomes of Example 2 was found to have a higher absorption rate than the control Gaba powder, a conventional natural liposome (Comparative Example 2).

In addition, as a result of measuring and comparing the concentration according to the intestinal absorption after 6 hours, the natural liposome of Example 2 was 82.4 mg / ml, and the control Gaba powder and the normal natural liposome (Comparative Example 2) were each 60.0 mg / ml, higher than that of 61.3mg / ml (Fig. 2b).

As shown in Figure 3a, the natural liposomes of Example 3 was found to have a higher absorption rate than the control Gaba powder (20%).

In addition, as a result of comparing the amount of change in the absorption rate of the membrane for 6 hours, the natural liposome of Example 3 was found to increase by 1.26, and it was found that the control group total increase was about 58% compared to the increase of 0.80 (Fig. 3b).

As shown in Figure 4a, the natural liposomes of Example 4 was found to have a higher absorption rate compared to 10% of the control group, Chongmyeongtang.

In addition, as a result of comparing the amount of change in the absorption rate of the membrane for 6 hours, the natural liposome of Example 4 was found to increase by 1.17 from 0.23 (0h) to 1.4 (6h), and 10% of the control group total myeongtang was 1.26 (6h) from 0.28 (0h). It was found to increase by 21% compared to the increase of 0.97 as (Fig. 4b).

[Experimental Example 3] MTT Assay Method

[Experimental Example 3-1] Measurement of absorbance according to concentration

Neuronal cells SH-SY5Y cells are seeded in 96 well plates at 1 X 10 ⁵ / mL, and after 24 hours have elapsed in DMEM medium (Dulbecco's modified eagle's medium, 10% FBS) at 37 degrees Celsius, 5 % CO2 conditions for 24 hours.

After incubation, H ₂ O ₂ was treated by concentration for 0 to 200 uM for 6 hours, and then maintained in MTT (1 mg / ml) solution for 4 hours. Thereafter, after dissolving using 0.04N HCl / isopropanol, the absorbance was measured at 570 nm using a spectrophotometer. The experimental results are shown in FIG. 5.

[Experimental Example 3-2] Measurement of cell protection effect according to each sample

0.1% of each sample in DMEM medium (Dulbecco's modified eagle's medium, 10% FBS) after 24 hours after seeding neuronal SH-SY5Y cells at 48 x 10 ⁵ / mL in a 48 well plate. And treated with H ₂ O ₂ at 25 uM at 37 ° C and 5% CO ₂ for 24 hours.

After incubation, H ₂ O ₂ was treated by concentration for 0 to 500 μM for 6 hours, and then maintained in MTT (1 mg / ml) solution for 4 hours. Thereafter, after dissolving using 0.04N HCl / isopropanol, the absorbance was measured at 570 nm using a spectrophotometer. The experimental results are shown in FIG. 6.

[Experiment result]

As a result of confirming the cell protective effect of barley fermentation-derived GABA against oxidative stress in brain neurons, as shown in FIG. 6, all samples showed higher cell viability than the control group treated only with H ₂ O ₂ .

When synthetic GABA (Sigma GABA) was used as a sample when the untreated Normal was set to 100%, 90% of fermented barley extract powder (65% of barley extract, 35% of barley yeast) was dissolved in deionized water as a sample. When used (barley fermentation-derived GABA), when using natural liposomes containing 5.7% of fermented barley extract powder 90% (barley extract 65%, barley yeast 35%) as a sample (barley fermentation-derived GABA natural liposomes), cell viability 89.14%, 82.90% and 78.45%, respectively.

In this way, when barley fermentation-derived GABA was simply natural liposomes (barley fermentation-derived GABA natural liposomes), it had a lower cell protective effect than synthetic GABA (Sigma GABA), but when it was treated with natural liposomes as in Example 2, it was simply natural liposomes. In one case, it was found that the protective effect of cells was higher than that of synthetic GABA.

[Experimental Example 3-3] Western Blot Method according to each sample-Gaba receptor and serotonin receptor activity of neurons confirmed

Neuronal cells SH-SY5Y cells are seeded in 6 well plates at 1 X 10 ⁵ / mL, and after 24 hours, Sigma GABA and fermented barley in DMEM medium (Dulbecco's modified eagle's medium, 10% FBS) GABA, fermented barley GABA natural liposomes, 0.1% of the sample of Example 2 were treated respectively, and after 3 days, the medium was changed and cultured again at 37 degrees Celsius and 5% CO2 for 6 days while processing the sample to differentiate.

After the completion of the culture, Western blot was performed to confirm the level of GANA receptor expression. After removing the medium, washed with PBS, and treated with 300uL of lysis buffer to prepare cell extracts, proceeded on a 10% SDS-Page gel and blocked for 1 hour using 5% skim milk. Thereafter, the primary antibody of the GABA Receptor protein was diluted 1000-fold and left overnight, then diluted with goat anti-rabbit horseradish peroxidase (secondary antibody) 5000-fold and reacted at room temperature for 3 hours.

Serotonin receptor and Gaba receptor proteins expressed on nitrocellulose membrane were photographed through X-ray film, and the results are shown in FIGS. 7 and 8.

[Experimental Example 4] Measurement of cell protection effect according to each sample

Neuronal cells SH-SY5Y cells are seeded into 48 well plates at 5 X 10 ⁵ / mL, and after 24 hours, normal, control, and donepezil in DMEM medium (Dulbecco's modified eagle's medium, 10% FBS) , Synthetic GABA, Chongmyeong-tang, Chongmyeong-tang natural liposome, 0.5% of the sample of Example 3 was treated with each%, and H ₂ O ₂ was cultured at 25 uM at 37 ° C and 5% CO2 for 24 hours.

After incubation, it was maintained in MTT (1 mg / ml) solution for 4 hours. Thereafter, after dissolving using 0.04N HCl / isopropanol, the absorbance was measured at 570 nm using a spectrophotometer. The experimental results are shown in FIG. 9.

As a result of confirming the cell protective effect of Chongmyeong-tang natural liposomes against oxidative stress in brain neurons, as shown in FIG. 9, all groups showed higher cell viability than the Control group treated only with H ₂ O ₂ . In particular, before the natural liposomes of Chongmyeongtang, the cell protective effect was lower than that of synthetic GABA (SIGMA GABA), but after the natural liposome treatment, the cell protective effect was higher than that of synthetic GABA. The natural liposomes of Chongmyeong-tang treated as in Example 3 were found to be particularly excellent in cell protection against oxidative stress.

Claims (18)

A liposome-constituent substance containing at least one natural emulsifier, a natural-derived solvent, a natural coating agent, and a natural preservative, wherein the liposome-constituent substance further contains sphingolipid, sterol and glycoproteins; And
Gaba, Baekmyeongtang, or mixtures thereof as a trapping material
It includes,
Based on the total weight of natural liposomes,
The content of the sphingolipids is 0.5 to 5% by weight,
The content of the sterol is 0.01 to 2% by weight,
The content of the glycoprotein is 2% by weight or more,
The sphingolipid: the sterol: the content ratio of the glycoprotein is 2: 0.1 to 0.5: 1 to 5 based on weight,
A natural liposome formed by mixing the natural emulsifier, the naturally-derived solvent and the gaba, cheonmyeongtang, or a mixture thereof by ultrasonic treatment. According to claim 1,
Based on the total weight of the natural liposome. 0.1 to 10% by weight of the natural emulsifier, 0.001 to 3% by weight of the natural coating agent, 0.001 to 3% by weight of the natural preservative, 0.1 to 30% by weight of the gaba, myeongtang, or mixtures thereof, the sphingolipids, sterols, sugar A natural liposome comprising 1.0-10% by weight of the total protein and the residual amount of the solvent derived from the natural. According to claim 1,
The natural emulsifier is at least one selected from the group consisting of lecithin, phosphatidyl choline, lysophosphatidyl choline, phosphatidyl ethanolamine, hydrogenated phosphatidyl choline, phosphatidic acid, phosphatidyl serine, phosphatidyl glycerol, phosphatidyl inositol and their hydrogenation products. Natural liposomes containing phospholipids. According to claim 1,
The natural emulsifier is a natural liposome comprising at least one fatty acid selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid. According to claim 1,
The natural-derived solvent is at least one natural liposome selected from the group consisting of distilled water, natural-derived glycerin, natural ethanol, natural propanediol, natural glycerin and fermentation alcohol. According to claim 1,
The natural coating agent is at least one natural liposome selected from the group consisting of maltodextrin, tapioca, guar gum, and corn starch. According to claim 1,
The natural preservative is a natural liposome that is one or more natural extracts selected from the group consisting of citrus extract, citron extract, eucalyptus extract, ginseng extract, clove extract, grapefruit extract. According to claim 1,
The sphingolipids are natural liposomes containing sphingomyelin. The method of claim 8,
The sphingomyelin is a natural liposome containing milk ceramide. According to claim 1,
The sterol is a natural liposome containing at least one sterol selected from the group consisting of phytosterol, campus terol, and vega pure. According to claim 1,
The sugar protein is a natural liposome containing carat collagen. delete delete According to claim 1,
Based on the total weight of natural liposomes,
The content of the sphingolipids is 1 to 3% by weight,
The content of sterol is 0.05 to 1.5% by weight,
The content of the glycoprotein is natural liposomes of 2.5% by weight or more. delete (a) a first step of mixing and sonicating a natural-derived solvent with at least one natural emulsifier at 35 to 80 ° C;
(b) a second step of mixing and ultrasonicating one or more selected from the group consisting of sphingolipids, sterols, and glycoproteins in the mixture of the first step;
(c) a third step of adding gaba, gongmyeongtang, or a mixture thereof to the mixture of the second step and ultrasonically mixing the mixture;
(d) a fourth step of mixing and sonicating the mixture of the third step with a natural coating at 35 to 80 ° C; And
(e) a fifth step of adding a naturally-derived solvent and a natural preservative to the mixture of the fourth step and sonication
Method of producing a natural liposome of claim 1 comprising a. A food composition comprising the natural liposome of claim 1 as an active ingredient. A food composition for improving learning and improving memory, comprising the natural liposome of claim 1 as an active ingredient.

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