KR20180129286A - Food composition for enhancing recognition ability comprising an fermented extract of aronia - Google Patents

Abstract

One embodiment of the present invention provides a food composition for enhancing recognition characterized by comprising an Aronia extract, a natural pigment, an excipient, and silicon dioxide having a concentration in a specific numerical range. Provided is the food composition for enhancing recognition which can provide expected improvement of dementia-related diseases having cognitive function improving effect in addition to the antioxidant ability of Aronia, which is known in the art, and which has an effect of inhibiting acetylcholinesterase and simultaneously increasing the expression level of BDNF, CREB and p-CREB due to the oral administration of the extract of Aronia within a specific concentration range in specific action mode.

Description

[0001] The present invention relates to a food composition for enhancing cognitive function,

The present invention relates to a cognitive function improving food composition comprising an extract of Aronia, and more particularly, to a functional health food comprising an Aronia extract having a cognitive function improving effect in a specific range of dosage .

Dementia is a syndrome that involves a wide range of mental functions. The incidence of dementia is estimated to be about 10% in people over 65 years old and 20% in people over 80 years old. The cause of dementia has been reported to be various, and other metabolic diseases of the brain such as Parkinson's disease, plasma disorder, and low frequency addictive or metabolic abnormalities are also causes of dementia. Alzheimer's disease begins with a decrease in memory, progressive memory cognitive impairment, and eventually a high degree of dementia. There are 10 million to 15 million patients worldwide, and the number is estimated to increase year by year. The pathological features of Alzheimer's disease are senile plaques and neurofibrillary tangles. In order to identify them, histological specimens of the brain must be observed histologically, and biopsy or autopsy must be performed. It is not practical in a clinical setting. Therefore, diagnosis is made based on the presence or absence of symptoms characteristic of Alzheimer's disease (memory, cognitive impairment, dementia).

Alzheimer's disease is an early manifestation of memory impairment and, as in the case of Parkinson's disease, patients with early stages of neurological deterioration are less likely to respond to pharmacotherapy. In addition, the rationale that choline drugs may help improve memory impairment has been reported (Corina Bejar, Eur J Pharmacol 383, 231-240, 1999). Thus, in the past, attempts have been made to improve the memory by using the Collin contract in Alzheimer's patients.

As a result, acetylcholine esterase inhibitors such as donepezil, galantamine and rivastigmine, which are currently used for improving memory disorders, have been used clinically but have low therapeutic efficiency and these synthetic drugs Vomiting, diarrhea, etc. Insomnia, insomnia, nightmare, muscle cramps, and other side effects that can cause the symptoms of temporary relief does not seem to be a fundamental solution. Recently, it is necessary to develop a therapeutic agent using natural materials having human compatibility.

Aronia melanocarpa (Black chokeberry), a perennial shrub, is a plant native to North America and northern Europe, which is used for edible or medicinal purposes and is also grown for ornamental purposes. The fruit of the Aronia tree belongs to berries containing anthocyanins such as strawberries, raspberries, cranberries, and blueberries. The fruit of Aronia has anthocyanin content of about 12 times that of cranberries, about 20 times of strawberries, It is known to be about four times as many as grapes and about 80 times as many as grapes. It is known that Aronia contains many kinds of anthocyanins such as Cyanidin-3-galactoside, Cyanidin-3-glucoside, Cyanidin-3-arabinoside and Delphinidin-3-glucoside and also contains a large amount of phenol. These studies have shown that not only do they have an excellent antioxidant effect, but also antiinflammation and immunity enhancement, as well as diabetes and various cardiovascular diseases, However, there is no report yet on the fact that Aronia has anti-aging and senile dementia

As the most common health functional beverage in the intake method, Aronia is the most problematic part when it comes to food, because people are reluctant to eat a lot because of their unique taste, bitterness and bitter taste. Specifically, it is believed that when the taste of an aroma extract health functional beverage is strong, the sour taste by the organic acid becomes strong first, and the tannin which is thought to be caused by the tannin and the dietary fiber as it goes back, I am reluctant to eat.

Therefore, it is necessary to develop functional capsicidal health food that can be easily ingested at any time and anywhere, while minimizing discomfort during eating and increasing the shelf life.

The present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a food composition comprising an astaxanthinum extract having an effect of improving cognitive function, while enhancing ease of feeding, as an active ingredient.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the precise form disclosed. There will be.

The present inventors have made intensive efforts to develop a food composition containing an optimum amount of an Aronia extract having a cognitive function improving effect while being able to easily ingest a savory taste of Aronia, To obtain a food composition containing crystalline cellulose and tapioca starch and the like.

As a technical means for achieving the above technical object,

According to an aspect of the present invention, there is provided a cognitive function improving food composition comprising an extract of Aronia, a natural pigment, an excipient, and silicon dioxide.

In an embodiment of the present invention, the atorvastatin extract may include 40 to 50 parts by weight based on 100 parts by weight of the total food composition.

In the embodiment of the present invention, the natural coloring matter may be any one selected from the group consisting of a cacao coloring, a gummy coloring matter, a gardenia yellow coloring, a safflowing yellow coloring, a bit red coloring, a purple sweet potato coloring, Which is a vegetable pigment.

In an embodiment of the present invention, the natural coloring matter may include 20 to 35 parts by weight based on 100 parts by weight of the total food composition.

In an embodiment of the present invention, the excipient may be any one selected from the group consisting of cellulose, lactose, starch, and combinations thereof.

In an embodiment of the present invention, the excipient may be 25 to 35 parts by weight based on 100 parts by weight of the total food composition.

In an embodiment of the present invention, the silicon dioxide may include 1 to 3 parts by weight based on 100 parts by weight of the entire health functional food.

In an embodiment of the present invention, the concentration of the atorvastatin extract may be 200 to 400 mg / kg.

In an embodiment of the present invention, the Aroma extract may be one that decreases the activity of acetylcholinesterase and at the same time increases neurotrophic factor (BDNF), CREB and p-CREB expression.

In an embodiment of the present invention, the effective daily dose of the food composition may be 1.0 to 1.5 g per day.

According to one embodiment of the present invention, there is provided a method for producing a food composition capable of maximizing a physiologically active substance such as anthocyanin contained in Aronia through a cognitive function improving food composition containing an extract of Aronia as an active ingredient and a stepwise process can do. In addition, the food composition of the present invention has an effect of inhibiting acetylcholinesterase activity, improving the activity of BDNF, CREB and p-CREB by improving the cognitive function by limiting the specific concentration of astaxanthin and the daily intake .

It should be understood that the effects of the present invention are not limited to the above effects and include all effects that can be deduced from the detailed description of the present invention or the configuration of the invention described in the claims.

Fig. 1 is a measurement of brain cervical cytoprotective activity in mouse hippocampus-derived cells.
Fig. 2 shows the measurement of the amount of intracellular reactive oxygen species in mouse hippocampal-derived cells.
Fig. 3 shows the amount of intracellular calcium ions measured in mouse hippocampus-derived cells.
Figure 4 shows the amount of glutathione measured in mouse hippocampal derived cells.
5 is a graph showing the amount of glutathione reducing agent measured in mouse hippocampus-derived cells.
6 shows the amount of glutathione peroxidase measured in mouse hippocampal-derived cells.
FIG. 7 shows the measurement of the 4-day escape time in an underwater maze experiment, which is an animal experiment related to cognitive function improvement.
Fig. 8 shows the measurement of the waiting time in the passive avoidance experiment, which is an animal experiment related to cognitive function improvement.
Fig. 9 shows the activity of acetylcholinesterase activity in mouse hippocampal-derived cells.
Fig. 10 shows the measurement of the amount of BDNF and CREB expression in mouse hippocampal-derived cells.
Fig. 11 shows the measurement of p-CREB expression in mouse hippocampus-derived cells.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. In order to clearly explain the present invention in the drawings, parts not related to the description are omitted.

Throughout the specification, when a part is referred to as being "connected" (connected, connected, coupled) with another part, it is not only the case where it is "directly connected" "Is included. Also, when an element is referred to as " comprising ", it means that it can include other elements, not excluding other elements unless specifically stated otherwise.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, the terms "comprises" or "having", etc., are used to specify that a feature, a number, a step, an operation, an element, or a combination thereof is described in the specification, , Steps, operations, elements, or combinations thereof, as a matter of principle, without departing from the spirit and scope of the invention.

Hereinafter, embodiments of the present invention will be described in detail.

In one aspect of the present invention, the cognitive function improving food composition comprises an atorvastatin extract, a natural pigment, an excipient, and silicon dioxide.

The Aronia of the above-mentioned Aronia extract may contain at least one of black chokeberry, red chokeberry, Aronia melanocarpaarone, Aronia melanoptera, Aronia melanoptera, and Aronia melanoptera. But is not limited thereto.

The method of extracting ammonia from the Aromania extract may further include at least one of an ultrasonic wave, an ultrahigh pressure, a high pressure pulse, a high pressure homogenization, or a freezing and thawing aerial process in a water extraction or alcohol extraction process such as ethanol or methanol. But is not limited to. More preferably, the aronia may be a 70% or more alcohol-extracted alcohol at room temperature for 80 to 100 hours. The above-mentioned alcohol can be a fermented alcohol produced by fermenting cereals with microorganisms or enzymes as a raw material, or a synthetic alcohol produced using ethylene as a raw material.

The extract of Aronia may contain 40 to 50 parts by weight, preferably 42.0 to 42.5 parts by weight based on 100 parts by weight of the total food composition. When the amount of the crude extract is less than 40 parts by weight in the total 100 parts by weight of the food composition, sufficient cognitive performance and antioxidative effect can not be obtained. When the amount is more than 50 parts by weight, tastiness due to auronic tannin and organic acid Can fall.

The natural pigment is produced not only from pigments contained in tissues and minerals of animals and plants but also by microorganisms produced by pulverizing, extracting and refining pigments, And preferably a vegetable coloring matter. The vegetable coloring matter may be at least one selected from the group consisting of a coloring matter, a high coloring matter, a marigold coloring matter, a berry coloring matter, a bit red, a turmerling coloring, an anthony coloring, an enamel coloring, a red cabbage coloring, Purple corn coloring, rutin, hibiscus coloring, safflower coloring, saffron coloring, cyan nut coloring, onion coloring, purple sweet potato coloring, purple coloring, purple coloring, purple coloring, purple coloring, The pigment may be a yam coloring matter, a chrysanthemum coloring matter, a carotene, a chlorophyll, a grape juice coloring matter, a pecan nut coloring, a Papia coloring, a tomato coloring, Safflower yellow pigment, bit red pigment, purple sweet potato pigment, grape skin pigment, and combinations thereof. The coloring and coloring density of the natural coloring matter changes depending on the environmental factors during the processing, storage and storage, and affects the merchantability and the sensibility of the processed food. The natural coloring is harmless to the human body and does not fade or discolor in the heating process Good heat resistance, good light resistance, chemical resistance. Natural pigments can be classified into carotenoids, flavonoids, pyrroles, and quinones, depending on the basic skeleton of the chemical structure. The natural pigments of the food composition may be carotenoids, flavonoids, or a combination thereof.

The natural coloring matter may include 20 to 35 parts by weight based on 100 parts by weight of the total food composition, preferably 12.0 to 18.0 parts by weight of a flavonoid coloring matter such as cacao coloring matter, carotenoid coloring matter 12.0 To 18.0 parts by weight. If the composition ratio of the natural pigment is less than 25 parts by weight, the characteristics of various bioactive components in the natural pigment may not sufficiently synergize with the active ingredient of the Aroma extract. If the composition contains more than 35 parts by weight, the weight ratio of the Aroma extract It can be expected that the cognitive performance improvement effect will be lowered.

The food composition may further comprise suitable excipients conventionally used in the manufacture. The excipient is difficult to fill the capsules by tableting with only the main ingredient when manufacturing the drug, and it serves to form the product. It is added for the purpose of giving a suitable hardness and shape to the medicine, or for the purpose of making it easy to handle with a certain capacity and weight when the amount of the main ingredient is small.

The form of the food composition may be granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions, Preferably a tablet or a capsule. For formulation into tablets or capsules, the active ingredient may further comprise an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Examples of excipients that may be included in the food composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, starch, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Lactose, methylhydroxybenzoate, talc, magnesium stearate and minerals, preferably starch, cellulose, lactose and their derivatives such as starch, cellulose, lactose, And combinations thereof. In the case of formulation, it is usually prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like. Such solid preparations are prepared by mixing at least one excipient or the like in the extract. In the case of the food composition, the excipient that can be added may be any one selected from the group consisting of cellulose, lactose, starch, and combinations thereof.

The cellulose of the excipient may be crystalline cellulose or microcrystalline cellulose, preferably crystalline cellulose. Microcrystalline cellulose Cellulose is a white, odorless, tasteless crystalline powder and a porous particle, which is a cellulosic which is purified and partially broken. Microcrystalline cellulose is used as a disintegrant for wetting and other tablets as excipients, suspending agents, excipients for tablets and capsules. Crystalline cellulose is purified by partially depolymerizing alpha-cellulose with mineral acid. It is composed of non-fibrous part and has fluidity. It is used as anti-caking agent, stabilizer, emulsifier or dietary fiber in oral administration.

The starch of the excipient is a tasteless white powder of powder and small particles, which can be used as an excipient, disintegrant and binder for fluidity improvement, tabletting and capsules. In addition to increasing the volume of the tablet, starch is also used as a binder and disintegrant, and a mixture with lactose is often used. All starches and flours may be suitable for use as starch and may be of any natural origin. It may also be a plant-derived starch obtained by standard mating techniques, including cross-breeding, translocation, inversion, transformation, or any other gene or chromosomal manipulation method including mutations thereof. Examples include potato starch, corn starch , Pregelatinized starch, pregelatinized starch, starchy potato starch, rice starch, tapioca starch or wheat starch, preferably tapioca starch. The starch may be 5 to 15 parts by weight, preferably 10 parts by weight, per 100 parts by weight of the total food composition.

The lactose of the excipient is white or whitish-colored particles or powder which is odorless and has a slight sweet taste. Lactose is mainly used for the purpose of increasing the volume of tablets. It is a stable substance, does not react with most drugs, is water-soluble, and has an advantage of quick release of liquor. The lactose may be alpha-lactose or beta-lactose and may be used in tablet and capsule excipients, lyophilization and infant formula.

In the present invention, silicon dioxide (SiO2) is distributed in the cell walls of diatoms or in sand or quartz. Silicon dioxide plays a role as an adsorbent capable of absorbing or adsorbing a liquid drug and making it into tablets, and it may be preferable to use colloidal silicon dioxide having high adsorption property and large specific surface area.

The silicon dioxide may include 1 to 3 parts by weight based on 100 parts by weight of the total food composition. If the amount is less than 1 part by weight, adsorption may be deteriorated and caking may not be prevented. If 3 parts by weight is exceeded, the silicon dioxide may be harmful to human body.

Improvement in cognitive function in the cognitive function improving food composition may mean a measure for preventing dementia such as Alzheimer's type dementia, cerebrovascular dementia, Pick's disease, Kruszfeld-Jakob disease, dementia caused by head injury, and Parkinson's disease.

There is no particular limitation on the type of food of the food composition. Examples of the food include dairy products including drinks, meat, sausages, bread, biscuits, rice cakes, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, various soups, beverages, alcoholic beverages, and vitamins And the like, and may include all health foods in a conventional sense.

The composition of the food composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, oral formulations, external preparation suppositories and sterilized injection solutions according to a conventional method, May be tablets, pills, powders, granules or capsules as solid dosage forms for oral administration.

In the present invention, the major component of the Aronia extract can be added directly to the food or can be used together with other food or food ingredients, and can be suitably used in accordance with conventional methods. The amount of the active ingredient to be mixed may be appropriately determined according to the purpose of use (for prevention or improvement). Generally, the amount of the extract in health food may be 0.01 to 50 parts by weight, preferably 40 to 100 parts by weight, 45 parts by weight. When the amount of the extract of Aronia is less than 40 parts by weight of 100 parts by weight of the whole food, the effect of improving the antioxidative and cognitive functions of the Aronia extract may be insignificant. When the amount of the extract is more than 45 parts by weight, And thus the palatability due to feeding can be lowered.

In the process for preparing the Aromania extract of the present invention, the Aroma fruit which has been washed with water and cleaned with the faeces was lyophilized, pulverized, and subjected to extraction at about 70% of the alcohol at room temperature for 96 hours, And concentrate. In the alcohol extracting step, pectin-decomposing complex bacteria or lactic acid bacteria may be added as needed, and ultrasonic irradiation can be performed intermittently.

In the food composition of the present invention, the above-mentioned Aromania extract is useful as an effective ingredient for the prevention and / or treatment of cardiovascular diseases, hypertension, or cognitive impairment due to the elimination of waste products in the body, toxin release, antioxidative effect, There may be an improvement effect. More specifically, the Ahnia extract has a protective effect against apoptosis, reduction of the amount of ROS which is an intracellular reactive oxygen species, cascade decrease of intracellular ROS due to decrease of intracellular calcium ion (Ca 2+) , An increase in the amount of glutathione, an intracellular antioxidant, an increase in the activity of the glutathione reducing agent, an intracellular antioxidant enzyme, an increase in glutathione peroxidase, an intracellular antioxidant enzyme, and a significant increase in the mitochondrial membrane potential . In relation to the activity of the extract of Aronia, in relation to the behavioral development using the animal test, the extract of Aronia with the concentration of 200-400 mg / kg was 94.5 ± 9% to 96.9 ± 7.5%, and the activity of acetylcholinesterase was significantly . And the above-mentioned extract of Aronia within the range of 200 to 400 mg / kg can simultaneously increase the activity of acetylcholinesterase as well as the expression of neurotrophic factor (BDNF), CREB and p-CREB.

The above-mentioned Aromania extract contains anthocyanins composed of cyanine, cyanidin, delpine, delpyridine, malvin, malvidin, pelargonin, pelargonidine, pheonin, pheonidine, petunin or petunidin. More specifically, the indicator component of the above-mentioned astaxanthin extract may be cyanidine-3-galactoside, and may be 1.00 to 2.00% by repeated extraction. The chemical formula of cyanidin 3-galactoside is shown in the following chemical formula 1.

Acetylcholinesterase is an enzyme that hydrolyzes acetylcholine to acetylcholine and choline. It acts on the parasympathetic nerves and the free ganglion, and removes synaptic signaling substances. Acetylcholine is one of the neurotransmitters of the autonomic nervous system, a basic neurotransmitter of the autonomic ganglion, acting on both the central nervous system and the peripheral nervous system and responsible for the cognitive function due to neurotransmission. Brain Derivated Neurotrophic Factor (BDNF) is a type of neurotrophic factor present in the brain. It is composed of 119 amino acids and a molecular weight of 12,3 kDa. It acts on neurons in both the central nervous system and peripheral nervous system. It has been known that efficacy has been shown in motor neuropathy model, drug toxicity, diabetes, traumatic microinvasive disorder model, retinal disorder model, Parkinson's disease model, Alzheimer's disease model and the like. The cAMP response element binding protein (CREB) is a cell transcription factor that binds to a specific binding DNA calling sequence and regulates translation of the downstream gene. Signals are activated through signal transduction pathways induced by binding to receptors. Activated CREB binds to the CRE region and invokes CBP (CREB binding protein), which regulates the activity of specific genes. CREB affects neuronal plasticity and long-term memory formation in the brain and is essential for spatial memory formation. CREB downregulation is associated with pathology of Alzheimer's disease, thereby increasing expression of CREB, which is being studied as a therapeutic target for Alzheimer's disease. P-CREB refers to the CREB protein whose activity is regulated by the binding of phosphate.

When the concentration of the extract is less than 200 mg / kg, the activity of neurofeedback (BDNF), CREB and p-CREB is not sufficient, and when the concentration of extract is more than 400 mg / kg The effect of reducing the activity of acetylcholinesterase may be deteriorated.

The preferred dosage of the composition of the present invention varies depending on the condition and body weight of the patient, the degree of disease, the drug form, the administration route and the period, but can be suitably selected by a person skilled in the art. However, in order to obtain the desired effect, it is preferable, but not limited, to administer the atorvastatin extract in the food composition at 0.5 to 1.5 g per day, preferably 1.0 to 1.5 g per day by oral administration. The above administration may be orally administered once a day orally or divided into several times, but preferably administered orally twice a day. The dose is not intended to limit the scope of the invention in any way.

The food composition of the present invention is not particularly limited as long as it contains the above-mentioned Aronia extract as an essential ingredient, and may contain various flavors or natural carbohydrates as an additional ingredient like ordinary food. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as dextrin, cyclodextrin, and the like

And sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .

In addition to the above-mentioned foods, the food of the present invention may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. These components may be used independently or in combination.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. For example, each component described as a single entity may be distributed and implemented, and components described as being distributed may also be implemented in a combined form.

The scope of the present invention is defined by the appended claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

Hereinafter, embodiments and test examples of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Example 1 . Aronia  Extract manufacturing process

Remove the faucet and prepare a 100 kg Arrhia freshwater bottle. 100 kg of freeze-dried Aronia was freeze-dried in a freeze dryer for 3 days, and the powder was pulverized into particles having a particle size of 1 to 10 μm through a high-speed pulverizing process and pulverized into 20 kg of lyophilized powder. The pulverized Aronia powder was immersed in 70% ethanol, and the arnoindereal components were extracted at room temperature for 96 hours. Then, the extract was filtered through a vacuum filter. Thereafter, the filtered extract was filtered and concentrated under reduced pressure at 40 ° C to remove the alcohol, followed by lyophilization and pulverization to obtain a powder.

Test Example 1 . Aronia  Measurement of surface components (high performance liquid chromatography HPLC ) Sample analysis)

An appropriate amount of an Aronia concentrate (hereinafter referred to as a standard solution) of Example 1 was dissolved in 100% methanol and dissolved to 1.0 mg / ml to prepare a standard stock solution.

The solution was dissolved by shaking, and diluted with methanol to prepare a standard solution. (E.g., 0.1, 0.05, 0.02, 0.01 mg / mL)

Approximately 0.5 g of the sample was taken, placed in a 100 ml volumetric flask, and adjusted to 70% methanol solution.

The solution was filtered through a membrane filter for filtration to obtain a test solution. The solution was analyzed by HPLC, UV detector and Waters symmetri C18 (4.6 mm ID x 250 mm, 5 μm) Or equivalent. HALO-RP (4.6 x 150 mm) column was used for HPLC analysis and methanol was used as an organic solvent for mobile phase. Separation of the isocratic was carried out up to 60 minutes. The detector was an Agilent 1100 series, and ultraviolet detection was performed at 520 nm. HPLC conditions and mobile phase conditions of the detection standard solutions are shown in Tables 2 and 3 below.

Item Condition Dose 10 μl Column temperature 30 ° C Mobile phase A solvent? Acetic acid: water = 1/9 (v / v)
B solvent? acetic acid: methanol: water = 10: 50: 40 Flow rate 1 mL / min Detector wavelength 520 nm

Time (minutes) menstruum A ( % ) B ( % ) 0 0 100 2 0 100 18 70 30 20 100 0 22 0 100 25 0 100

Test Example 2 . Aronia  Extract Neuronal cell  Measure protective activity

In order to confirm the improvement of cognitive function of the extract of Aronia, the protective activity test of brain neurons was carried out. HT22 cells derived from mouse hippocampus were used. HT22 cells were cultured in DMEM supplemented with 10% FBS in a 5% CO 2 incubator at 37 ° C. The HT22 cells were treated with negative control, glutamate alone, 50 mg / ml of glutamate + astrocyte extract, 100 mg / ml of glutamate + astrocyte extract and 200 mg / ml of glutamate + The results are shown in FIG. 1 below.

As a result of the measurement, the cell survival rate of 86.49 ± 2.77% at a concentration of 200 mg / ml in the group treated with Aronia showed that the cells were protected in a concentration-dependent manner.

Test Example 3 . In HT22 cells ROS amount  Measure

After 5 groups were prepared in the same manner as in Test Example 2, the amount of ROS, which is an intracellular reactive oxygen species, was measured. The results are shown in FIG.

In the measurement results, in the group treated with Aronia, the ROS amount was 104.40 ± 12.39% at the concentration of 100 mg / ml and 87.54 ± 12.82% of the ROS amount at the concentration of 200 mg / ml, Respectively.

Test Example 4 . In HT22 cells Ca2 + Measurement

After 5 groups were prepared in the same manner as in Test Example 2, the amount of intracellular calcium ions (Ca < 2 + >) was measured. Intracellular calcium ion induces oxidative stress by increasing the amount of ROS in the cells.

As a result of the measurement, the amount of calcium ion in the group treated with Aronia was 113.33 ± 11.60% at a concentration of 200 mg / ml, and the amount of calcium ion was decreased in a concentration-dependent manner.

Test Example 5 . In HT22 cells Amount of glutathione  Measure

After preparing five groups as in Test Example 2, the amount of glutathione was measured. The results are shown in FIG. 4 below.

As a result of the measurement, glutathione levels of 44.85 ± 5.98% at 200 mg / ml of the group treated with Aronia showed a significant increase in the amount of glutathione.

Test Example 6 . In HT22 cells Glutathione  Reducing agent and Amount of peroxide  Measure

After 5 groups were prepared in the same manner as in Test Example 2, the amount of glutathione reducing agent and peroxide was measured. The results are shown in FIGS. 5 and 6. Glutathione reductase and peroxidase reduce glutathione and reduce intracellular oxidative stress.

The results showed that the concentration of glutathione reductant and peroxidase in the group treated with Aronia was 32.31 ± 3.85% and 67.41 ± 11.15% at the concentration of 200mg / ml, respectively, Respectively.

Test Example 7 . Aronia  Experimental measurement of underwater labyrinth of fermented extract

i) Preparation of experimental animals

Four-week-old SPF male ICR mice (BioLink, Chungbuk, Korea, Korea) were used in this experiment. After the acquisition, the adaptation period in the breeding environment was completed for one week. The animals were weighed during the adaptation period, and the selected animals were distributed uniformly to the average weight of each group. The animals were given a temperature of 22 ± 2 ℃, a relative humidity of 50 ± 10%, a ventilation frequency of 10 to 15 times / hr, 12 hour period (09: 00 ~ 21: 00), and adapted environment conditions were set for 7 days in a dark / light breeding room. During the adaptation period, feed and water were supplied free of limitations, and seven animals were made per experimental group.

Each experimental group was divided into the control group, the scopolamine group which is a memory decaying substance, the donepezil group as a positive control group, the donor group as a preventive substance for memory decline, the aroinia group as a concentration, and the anthocyanin group as a concentration.

ii) Measurement of underwater labyrinth experiment

Fill the water maze pool (diameter: 90 cm, height: 45 cm) with water mixed with milk (water temperature: 20 ± 1 ° C) according to Morris's method. The pool was divided into four parts and a white platform (diameter: 6 cm, height: 29 cm) was placed in the center and the platform was immersed about 1 cm to make it invisible on the surface of the water.

For adaptation, the first day of the experiment was performed without swimming platform for 60 seconds, and from the second day, the test trial was performed for 4 days in the pool where the platform was placed. The time for escape latency Respectively. If the mouse does not escape to the platform within 120 seconds, it is raised directly above the platform, allowing it to escape for 10 seconds. Position the platform for 4 days and position the mouse differently.

The scopolamine (1 mg / kg) was dissolved in physiological saline and injected subcutaneously 30 minutes before the training. The scopolamine extract (50, (1, 5, 10, 25 mg / kg), cyanidin-3-O-galactoside + cyanidin- 3-O-glucoside (10 mg / kg) and donepezil (1 mg / kg), a positive control, were administered orally to determine the effect of scopolamine on dementia induced by scopolamine.

The results are shown in FIG. 7. As shown in FIG. 7, the improvement activity of scopolamine-induced memory impairment in all the experimental groups was confirmed by an underwater maze test. As a result, the escape latency s) and lowest escape latency (s) (53.6 ± 5.3 s and 54.9 ± 0.8 s) at concentrations of 400 and 600 mg / kg, respectively. But not significantly at concentrations below 200 mg / kg. In the case of cyanidin-3-O-galactoside, cyanidin-3-O-galactoside + cyanidin-3-O-glucoside (10 mg / kg) decreased escape lantency Did not reduce the escapel latency (s) increased by scopolamine. It was found that escape latency (time to visit target in water) gradually decreased during 4 days of experiment.

Test Example 8 . Aronia  Passive avoidance experiment measurement of fermented extract

i) Preparation of experimental animals

An experimental animal was prepared in the same manner as in Test Example 7 above.

ii) Measurement of passive avoidance experiment

The manual avoidance measuring device is divided into a light box and a dark box (40 x 20 x 20 cm), and a door is provided between the rooms to allow movement of the mouse. A mouse placed in a light box with a 3 mm thick stainless steel rod at 0.5 cm intervals on the bottom of the box was given an electrical stimulation of 0.1 mA / 10 g body weight through a stainless steel rod when it entered the dark box. After 24 hours, the same test is performed to measure the time that the mouse stays in the bright room, and it is used as an index to memorize the training of the day before. Samples were administered orally and scopolamine was subcutaneously administered.

Anthocyanin A (1, 5, 10, 25 mg / kg), anthocyanin A + B (10 mg / kg) and a positive control group (50, 100, 200, 400 and 600 mg / kg) Donepezil (1 mg / kg) was orally administered. After 90 minutes, the control group was subcutaneously administered the normal saline and scopolamine (1 mg / kg) was subcutaneously administered to the other groups. Scopolamine was administered 30 minutes later and the experiment was carried out. After 24 hours, the same experiment was carried out and the time at which the mouse moved to the dodge box was measured by escape latency time. If there was no movement for 180 seconds, the experiment was stopped.

The results are shown in FIG. 8. As shown in FIG. 8, the improvement activity of scopolamine-induced memory impairment in all experimental groups was confirmed by passive avoidance experiment, and the latency time decreased by scopolamine in a concentration-dependent manner The highest latency time (131.0 ± A 14.8 s) was found in 400 mg / kg of the Ahnonia extract and the latency time was reduced to 400 mg / kg at the concentration of 600 mg / kg. Anthocyanin A (cyanidin-3-O-galactoside) also increased the latency time reduced by scopolamine. Cyanidin-3-O-galactoside + cyanidin-3-O-glucoside (10 mg / kg) did not show cognitive function improving activity.

Test Example 9 . In HT22 cells Acetylcholinesterase  Inhibition measurement

In order to confirm the activity of improving the cognitive function of the extract of Aronia, the hippocampus was extracted from the mouse after the animal test, and the acetylcholinesterase activity was measured.

The results are shown in FIG. 9. More specifically, in the case of Scopolamine-treated mice, acetylcholinesterase activity of 120,9 ± 23% was measured and increased compared to the control group. In the case of astaxanthin extracts, the concentrations of acetylcholinesterase of 113.2 ± 4.0%, 107.0 ± 15.4%, 94.5 ± 8.8%, 96.9 ± 7.3%, and 107.8 ± 2.1%, respectively, at concentrations of 50, 100 200 400 and 600 mg / Activity, and the activity of acetylcholinesterase increased by scopolamine was significantly decreased, but it was found to be slightly increased at the concentration of 600 mg / kg.

Test Example 10 . Neurotrophic Factors in HT22 Cells BDNF ), CREB  And p- Of CREB  Expression measurement

In order to confirm the mechanism of activity of improving the cognitive function of the extract of Aronia, the hippocampus was extracted from the remaining mice except donepezil-treated mice, which were positive control group after the animal test, and the amount of BDNF, CREB and p-CREB expression was measured .

The results are shown in FIG. 10 and FIG. 11. More specifically, the scopolamine-treated mice exhibited lower expression values (0.83 +/- 0.05) than the control group BDNF expression. The amount of BDNF, a neurotrophic factor reduced by scopolamine, was significantly activated in the hippocampus of the mice treated with the allnia extract at all concentrations. CREB, a regulator of BDNF expression, expressed 0.84 ± 0.06 in scopolamine - treated mice. It was confirmed that p-CREB was significantly activated in the extract of Aronia.

Example 2 . Aronia  Preparation of food composition containing extract

The capsule composition was prepared as a food composition containing the extract of Aronia extracted according to Example 1 as a main active ingredient. The composition ratio of the food composition is shown in Table 1 below.

Raw material name content(%) Aromania extract 42.3 Crystalline cellulose 17.3 Cacao pigment 14.2 Gardenia pigment - A 14.2 Tapioca starch 10.0 Silicon dioxide 2.0

Claims (10)

Aronia extract;
Natural color;
Excipients; And
Silicon dioxide;
Wherein the cognitive function improving food composition is a cognitive function improving food composition. The method according to claim 1,
Wherein the atorvastatin extract comprises 40 to 50 parts by weight based on 100 parts by weight of the total food composition. The method according to claim 1,
Wherein the natural coloring matter is a vegetable coloring matter selected from the group consisting of a cacao coloring matter, a gummy coloring matter, a gardenia yellow coloring, a safflowing yellow coloring, a bit red coloring, a purple sweet potato coloring, a grape skin coloring and a combination thereof Cognitive performance improving food composition. The method according to claim 1,
Wherein the natural coloring matter comprises 25 to 35 parts by weight based on 100 parts by weight of the total food composition. The method according to claim 1,
Wherein the excipient is any one selected from the group consisting of cellulose, lactose, starch, and combinations thereof. The method according to claim 1,
Wherein said excipient comprises 25 to 35 parts by weight of said excipient based on 100 parts by weight of the total food composition. The method according to claim 1,
Wherein the silicon dioxide comprises 1 to 3 parts by weight based on 100 parts by weight of the whole health functional food. The method according to claim 1,
Wherein the concentration of the atorvastatin extract is 200 to 400 mg / kg. The method according to claim 1,
Wherein said atorvastatin extract reduces the activity of acetylcholinesterase and simultaneously increases the expression of neurotrophic factor (BDNF), CREB and p-CREB. The method according to claim 1,
Wherein the effective dose of the food composition is 1.0 to 1.5 g per day

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