WO2012036506A2

WO2012036506A2 - Composition for promoting memory and learning ability

Info

Publication number: WO2012036506A2
Application number: PCT/KR2011/006853
Authority: WO
Inventors: 이창호; 김인호; 한대석; 김영언; 박동준; 최준혁; 이혜성; 조승목
Original assignee: 한국식품연구원
Priority date: 2010-09-17
Filing date: 2011-09-16
Publication date: 2012-03-22
Also published as: WO2012036506A3

Abstract

The present invention relates to a composition for promoting memory or learning ability and to a composition for preventing and treating cognitive impairment, which comprise one, two or more plant extracts selected from the group consisting of Artemisia apiaceae extract, Illicium verum extract and Lepidium apefalum extract. The present invention has the effect of inhibiting neural cell damage, especially, neural cells of the basal part of the cerebrum by inhibiting acetylcholinesterase activity and through antioxidative activity (for example, inhibiting the generation of oxidative reactive species) and affinity for an NMDA receptor. The compositions of the present invention may have the effect of not only promoting memory or learning ability by protecting neural cells and preventing damage to neural cells but also of preventing and treating diseases caused by cognitive impairment. In addition, according to the present invention, Lepidium apefalum extract is provided as basic material for pharmaceuticals or food which has efficacy in promoting memory or learning ability or efficacy for preventing and treating cognitive impairment.

Description

【Specification】

[Name of invention]

Memory and Learning Competency Composition

[Technical field]

The present invention relates to a composition for enhancing memory and learning ability, comprising chungho extract, octagonal fennel extract or sperm extract as an active ingredient.

[Background technology]

There are 100 billion neurons in the human brain, each of which is connected to other nerve cells around each other by an average of 10,000 synapses, forming a network. Through these synapses, the close communication between the neurons is achieved, and at this time, a mediator is a neurotransmitter (neurotransmitter). Neurons release neurotransmitters to stimulate receptors and thereby exchange information with each other. The way that the human brain stores memory is the process of strengthening synaptic connection strength through continuous chemical and electrical stimulation, which is called long-term potentiation (LPT). The characteristics of synapses that are enhanced is called synaptic plasticity. The fact that neurotransmitters chemically communicate information at synapses was experimentally proven at the beginning of the 20th century. Currently known neurotransmitters include acetylcholine (ACh), norepinephrine, dopamine (DA), gaba (GABA), glycine and glutamic acid (Table 1). Table 1

Gaseous nitric oxide intestinal, midbrain expansion, memory nerve cell death

(NO) boundary (cerebellum (nitroglycerin), etc.)

Cognitive impairment, such as the loss of memory and learning ability caused by Alzheimer's disease, a type of senile dementia, is caused by damage to acetylcholine neurons in the base of the cerebral base, and according to a study on the muscarinic acetylcholine receptor Agonists, acetylcholine production promoters, and acetylcholinesterase inhibitors have been developed to enhance the function of acetylcholine neurons according to various mechanisms of action. Currently, tacrine has been developed as an acetylcholinesterase inhibitor, but there are some side effects such as gastrointestinal disorder. Therefore, it is necessary to search and utilize inhibitory active material from natural plant material.

Cheongho lrtemisia apiaceae Herbal is a herbaceous herb that is a biennial herb that is commonly found in the subterranean field of Korea, in the middle of Korea, or on the sandy ground near the stream. 40-150 cm in size, without hairs all over, with many branches on the stem. Its origin is the outpost of the wormwood, which is collected before summer flowers bloom and dried in the shade. The main ingredients are abotamine, vitamin A, β-bourbonne, β-bourbonene, caryophyl lene, α-pinene, β-pinene, 1 , 8-cineol, and α-thujone. Efficacy is known as hemostasis, fever, edema, streptococcal effect.

The octagonal verum Hook /.) Is a fruit of the lysium burum, belonging to the magnolia family, and grows in tropical Asia. Staranis is named for star fruit, and chewing a small amount after a meal promotes digestion. One with fever, 1-2 cm long, 0.3-0.5 in diameter. The outside is reddish brown with irregular wrinkles. The interior is pale brown, soft and shiny. The stem of the fruit is 3-4 cm long and connected to the lower part. Seeds in the fruit are 6 길이 long and are reddish brown or yellowish. It is brown. It is fragrant and tastes sweet and irritating. The fruit contains about 5% of essential oils and 80-90% of the essential oils are aneul. In addition, pinene, fellandene, cineol, limonene, dipentene, vesicles, aniseketone, anisealdehyde, aniseic acid.

Energized by the dam, Lepidium apefalum f /, is found in Korea, Central Asia, the Himalayas, and China. It is native to North America and grows in the clearings around fields or licensees. Seeds are small brown discoid, with white membranous wings at the edge. Energizers include fatty oils, Sinigrins and sugars. Energized as a herbal seed is used for cough, asthma, pulmonary tuberculosis, exudative pleurisy, swelling and poor urination, and heart failure.

Flavonoids that have a positive effect on the human brain include isoflavones found in soybeans, flavanols found in tea and cocoa, red wine, and anthocyanins. Silver isoflavones mimic the action of estrogens in the brain, while flavanols and ansocyanins mitigen-activated protein kinase (MAPK) pathways and PI3 kinase (PI3 kinase) / Akt signaling Transduction Stages Interact with signaling pathways in neurons such as reactions. Both MAPK and PI3 kinase pathways are known to have memory storage functions in the hippocampus and cortex of the brain. It has the potential to enhance memory and learning by activating these pathway kinases. It is also known to promote the activation of proteins such as cAMP response element binding (CREB), which is involved in the expression of genes associated with memory. Research teams at Kings College London in the UK have found that ingredients in broccoli, potatoes, oranges, apples and radishes work the same way as Alzheimer's treatments, which are neurotransmitters. It has been shown to inhibit acetylcholinesterase, an enzyme that destroys phosphorus acetylcholine.

Therefore, agonists and antagonists capable of promoting acetylcholinesterase inhibitors and receptor activities from herbal and plant materials, and proteins related to improving memory and cognitive function There is a need for a substance that can search for activating functions and use selected natural plant materials to treat and prevent memory and cognitive impairment. Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

[Detailed Description of the Invention]

[Technical problem]

The present inventors made an effort to develop a material that is safe for the human body, especially a plant-derived material, which can effectively enhance memory or learning ability, and as a result, the blue-green, octagonal fennel and energetic energy that has been conventionally used as a herbal medicine has a memory or learning ability. The present invention has been completed by proving that it is very effective in promoting this.

Therefore, an object of the present invention is to provide a food composition for improving memory or learning ability comprising one or two or more plant extracts selected from the group consisting of green leaf extract, octagonal fennel extract and sperm extract as an active ingredient.

Another object of the present invention to provide a pharmaceutical composition for preventing or treating cognitive dysfunction comprising one or two or more plant extracts selected from the group consisting of green leaf extract, octagonal fennel extract and sperm extract as an active ingredient.

Another object of the present invention to provide a food composition for improving cognitive dysfunction comprising one or two or more plant extracts selected from the group consisting of green leaf extract, octagonal fennel extract and sperm extract as an active ingredient. Other objects and advantages of the present invention are described in the following detailed description, The claims and drawings are more apparent.

Technical Solution

According to an aspect of the present invention, the present invention provides a food composition for improving memory or learning ability comprising one or two or more plant extracts selected from the group consisting of green leaf extract, octagonal fennel extract and sperm extract as an active ingredient. .

According to another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cognitive dysfunction comprising one or two or more plant extracts selected from the group consisting of chungho extract, octagonal fennel extract and sperm extract as an active ingredient. to provide.

According to another aspect of the invention, the present invention provides a food composition for improving cognitive dysfunction comprising one or two or more plant extracts selected from the group consisting of chungho extract, octagonal fennel extract and sperm extract as an active ingredient. .

The present inventors made an effort to develop a material that is safe for the human body, especially a plant-derived material, which can effectively improve memory or learning ability, and as a result, the Chungho, Octagonal Fennel and Energizer, which have been conventionally used as a herbal medicine, have a memory or learning ability. It was found to be very effective in promoting the When the plant extract used in the composition of the present invention is obtained by treating the extractant with a plant, various extractants may be used. Preferably, a polar solvent or a nonpolar solvent can be used. Suitable polar solvents include (i) water, (ii) alcohols (preferably methanol, ethane, propanol, butanol, normal-propane, iso-propane, normal-butane, 1-pentanol, 2 -Butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) D FOCdimethyl- formamide) and (v) dimethyl sulfoxide (DMS0). Suitable as nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, nucleic acid, 2,2, 4-trimethylpentane, decane, Cyclonucleic acid, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane 1-chloropentane, 0-xylene, diisopropyl ether, 2-chloropropane, toluene, 1—chloropropane, chlorobenzene, Benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, anneal, diethylamine, ether, carbon tetrachloride and THF.

More preferably, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propane, butane, etc.), (c) the lower alcohol Mixed solvent of water with (d) acetone, (e) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) nucleic acid and (j) diethyl Ether. Most preferably, the extract of the present invention is obtained by treating water, methane, ethane or a combination thereof in clarity.

As used herein, the term 'extract' has the meaning commonly used as a crude extract in the art as described above, but also broadly includes a fraction additionally extracting the extract. That is, the plant extract of the present invention includes not only those obtained by using the aforementioned extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation column by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), and various additionally performed The fraction obtained through the purification method is also included in the plant extract of the present invention.

The plant extract used in the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze drying or spray drying. The composition of the present invention works very effectively in preventing and treating cognitive impairment as well as enhancing memory or learning ability.

The effects of the present invention are exerted by inhibiting the protection and damage of neurons, preferably acetylcholinergic neurons in the base of the cerebral base.

Acetylcholine is a neurotransmitter that is secreted at the nerve endings and delivers nerve stimuli to muscles. After stimulation is completed, acetylcholinesterase is used to Decomposes to acetate.

According to a preferred embodiment of the present invention, the present invention can inhibit the degradation of acetylcholine by acetylcholinesterase by inhibiting the activity of acetylcholinesterase can inhibit the protection and damage of nerve cells.

In addition, the present invention not only inhibits memory loss and learning ability degradation due to neuronal damage through antioxidant activity, but also prevents neuronal cell damage and exerts an effect of improving memory or learning ability.

As used herein, the term "active oxygen species" means that oxygen, which is essential for organisms that breathe organically, is incompletely reduced during enzymatic and most electron transport or energy metabolism processes, or peptide growth factors, cytokines. And oxygen by-products generated by stimulation of various actions. In other words, reactive oxygen species have a free radical (a form in which an atom or molecule has electrons unpaired in its outer orbit), which means that it is unstable and thus has strong activity.

Reactive oxygen species are sometimes referred to as harmful oxygen because excessive production leads to toxicity to the living organism, that is, oxidative stress. By oxidizing huge molecules (proteins, lipids, etc.) in the cell, it destroys the homeostasis of the cells, kills the cells, and causes fatal damage in the tissues of the cells, cancer, dysplasia, Alzheimer's disease, Parkinson's disease, ischemic diseases This is because it is related to the causes of various degenerative diseases such as atherosclerosis and general aging.

For example, reactive oxygen species include lipid peroxide (lipid) in addition to superoxide anion radical (0 ^{2 "} ), hydrogen peroxide (H ₂ 0 ₂ ), and hydroxy radical (0H). peroxide), nitric oxide (NO), peroxynitrite (peroxinitrite: N0 ₃ ^2_ ), thiol peroxy radical (R-S0 ^2_ ), and ¾0 ₂ is TGF, -1, which is produced by stimulation such as PDGF and EGF, attracts most attention as a secondary signaling substance because it is relatively stable, less toxic, and easily diffuses to the cell membrane unless it reacts with transition metals. Because it can pass through This is because it has suitable properties as a signaling material that must be produced and destroyed quickly.

According to a preferred embodiment of the present invention, the present invention has an antioxidant activity, more preferably inhibits or eliminates the generation of reactive oxygen species, and ultimately inhibits the damage of neurons to memory or learning ability It is a composition capable of preventing and treating cognitive dysfunction as well as enhancement.

The present invention binds to N-methyl-D-aspartate (NMDA) receptors to inhibit neuronal cell death, thereby inhibiting neuronal cell protection and damage. Therefore, the present invention can prevent and treat cognitive impairment as well as inhibit the decrease in memory or learning ability due to neuronal damage in combination with the NMDA receptor.

According to a preferred embodiment of the present invention, the present invention acts as an antagonist of an N-methyl-D-aspartate (NMDA) receptor to inhibit neuronal damage, thereby improving memory or learning ability and preventing cognitive impairment. And treatment.

The term 'cognitive impairment' in the specification of the present invention refers to a disease that does not exhibit cognitive functions such as memory processing, perception and problem solving.

The present invention is a composition capable of preventing and / or treating cognitive dysfunction. More specifically, the present invention is acetylcholinesterase

It is a composition that can inhibit and / or treat cognitive dysfunction by inhibiting the activity of (Acetylcholinesterase), antioxidant activity (eg reactive oxygen species) and NMDA (N-methyl-D-aspartate) receptor.

According to a preferred embodiment of the present invention, the cognitive impairment in the present invention is dementia, learning disorder, agnosia, amnesia, aphasia, apraxia or Delirium, more preferably AIDS-induced dementia (AIDS dementia com lex), Binswanger's disease, dementia with Lewy Bodies, frontotemporal dementia, mildness Cognitive impairment, multi-infarct dementia, pick's disease, semantic dementia dementia), Alzheimer's dementia or vascular dementia. The composition of the present invention may be prepared as a pharmaceutical composition.

According to a preferred embodiment of the present invention, the composition of the present invention comprises (a) a pharmaceutically effective amount of the plant extract of the present invention described above; And (b) a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve the efficacy or activity of the above-mentioned clear green extract.

When the composition of the present invention is made into a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation of lactose, dextrose, sucrose, sorbbi, manny starch, acacia gum, phosphate chamomile, alginate, gelatin , Calcium silicate, microcrystalline cellulose, polyvinylpyridone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil One is not limited thereto. The pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and preferably applied by oral administration.

Appropriate dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as formulation method, mode of administration, age of patient, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and reaction response. Can be. Typical dosages of the pharmaceutical compositions of the invention are in the range of 0.001-100 mg / kg on an adult basis.

The pharmaceutical composition of the present invention may be formulated using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those skilled in the art. It may be prepared in unit dose form or incorporated into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oil or aqueous media, or may be in the form of axes, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers. The composition of the present invention may be provided as a food composition.

When the composition of the present invention is made of a food composition, as an active ingredient, as well as green leaf, octagonal fennel or sperm extract, it contains the components commonly added in the manufacture of food, for example, protein, carbohydrate, fatty nutrients , Seasonings and flavorings. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugar alcohols such as polysaccharides such as conventional sugars such as dextrin, cyclodextrin, and xyl, sorbitol, erythritol and the like. As the flavoring agent, natural flavoring agents [tautin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared with a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, jujube extract, jujube extract, licorice extract, and the like may be further included. . According to another aspect of the present invention, the present invention comprises administering one or more plant extracts selected from the group consisting of Cheongho Artemisia apiaceae extract, Octagonal Ullicium verum) extract and Energetic Lepidhim apefalum extract Provide ways to improve memory or learning skills. According to another aspect of the invention, the invention comprises the step of administering one or more plant extracts selected from the group consisting of Cheongho Artemisia apiaceae) extract, Octagonal UUici verum) extract and Energetic iLepidi apefalum) extract Provides methods for preventing or treating cognitive disorders. Since the plant extract used in the present invention has been described above, the description thereof is omitted to avoid excessive duplication.

Advantageous Effects

The features and advantages of the present invention are summarized as follows:

(i) The present invention comprises a composition for improving memory or learning ability and a composition for preventing and treating cognitive dysfunction, comprising one or two or more plant extracts selected from the group consisting of green leaf extract, octagonal fennel extract and sperm extract as an active ingredient. To provide.

(ii) The present invention has the effect of inhibiting the activity of acetylcholinesterase, antioxidant activity (eg, reactive oxygen species) and affinity with NMDA receptors to inhibit the damage of nerve cells, especially nerve cells at the base of the cerebrum. Have

(iii) The present invention not only enhances memory or learning ability through protecting and preventing neurons, but also exerts effects of preventing and treating diseases with cognitive impairment.

(iv) The present invention also provides basic data as medicines and foods of plant extracts having the effect of enhancing memory or learning ability, or preventing and treating cognitive impairment.

[Brief Description of Drawings]

1 is a diagram showing the AChE inhibitory effect of chungho ethane extract of the present invention (Fig. La), octagonal ethane extract (Fig. Lb) and energetic ethanol extract (Fig. Lc).

Figure 2 is a result showing the antioxidant activity of the clarified ethanol extract (Fig. 2a), octagonal fennel ethanol extract (Fig. 2b) and energetic ethanol extract (Fig. 2c) of the present invention.

Figure 3 is a result showing the NMDA receptor binding effect of the clarity ethanol extract (Fig. 3a), octagonal fennel ethanol extract (Fig. 3b) and energetic ethanol extract (Fig. 3c) of the present invention. Figure 4 is a diagram showing the improvement effect of the ethanol extract in the scopolamine-induced memory deficiency rats in the step-through latency (STL) to avoid the light to enter the dark room during the passive avoidance experiment.

Figure 5 is a diagram showing the effect of chungho ethanol extract on the memory of scopolamine-treated rats in a radial eight-way maze experiment.

[Form for implementation of invention]

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples according to the gist of the present invention to those skilled in the art to which the present invention pertains. Will be self explanatory. Example

Throughout this specification, unless otherwise indicated, "" used to indicate the concentration of a particular substance is solid / solid (weight / weight)%, solid / liquid (weight / volume), and liquid / Liquid is (volume / volume)%. Experimental Materials and Methods

Cheongho used in the experiment was purchased from Jecheon Oriental Herbal Medicine (http://www.jchanbang.com) site and dried in the blended KA2610 (Jworldtech, South Korea). 80% ethanol was added to 2 kg of pulverized clarity powder at a ratio of 1:10 and extracted at 95 ^° C. for 6 hours based on the time of boiling at the C0SM0S-660 ultra-fast vacuum cold extractor (kyungseo, South Korea). Chungho extract was concentrated under reduced pressure at 80 ^° C and used as a test sample.

Octagonal fennel and sperm were purchased from Dongin Herbal Pharmaceuticals (China). 80% ethanol was added to the dried octagonal fennel and sperm, respectively, and extracted and concentrated at 95 ° C for 6 hours using a C0SM0S-660 ultrafast vacuum cold extractor. Concentrated Samples were stored at -80 ° C and frozen and dried with a lyophilizer to use powdery samples. The sample was dissolved in tertiary distilled water or DMS0 (dimethyl sulfoxide) immediately before the experiment to prepare a sample sample. Measurement of AChE (Acetylcholinesterase) Inhibitory Activity

Determination of AChE (Acetylcholinesterase) inhibitory activity of chungho extract, octagonal fennel extract and sperm extract was used by modifying the method of Ellman et al. (Ellman, GL, et al, Biochem. Pharmacol., 7: 88-95, (1961) )). AChE inhibitory activity is yellow due to the reaction product of thiocholine, which is produced when AChE hydrolyzes acetylthiochol ine, with DTNB (5,5'-dithio bis (-2-nitrobenzoic acid)). Since NTB (2-nitrobenzoic acid) is produced, the resulting NTB was observed by measuring the absorbance at 412 ran.

That is, 500 ≠ a cuvette is added as a semi-aperture, 0.1 M SPB (sodium phosphate buffer; pH 8.0) 445 ^ and sample sample solution 25 ^ (final concentration 100; «g /) are added, and the coupling agent 0.001 M DTNB 18 / ^ (Sigma Aldrich, St. Louis, MO, USA) and 0.0075 M acetylthiocholine iodine 6 μί (Sigma Aldrich, St. Louis, MO, USA) were added and reacted for 4 minutes in a 25 ^° C thermostat. . After reaction, the enzyme Aceticocholinesterase 6 ≠ (0.072 unit, Sigma Aldrich, St. Louis, M0, USA) was added and reacted for 12 minutes in a 25 ^° C incubator, and then measured the light intensity at 412 nm. .

All experiment groups were repeated three times to obtain the average value of absorbance, AChE inhibitory effect of the extract of the present invention is the absorbance value of the control group in the same amount of 0.1 M SPB (pH 8.0) instead of the extract of the present invention and the present invention Using the absorbance value of the experimental group containing the extract was calculated as follows. On the other hand, takrine (Sigma Aldrich, St. Louis, MO, USA) and Eserine (Sigma Aldrich, St. Louis, MO, USA) at 100 μg / m ^ concentration were used as a positive control. Inhibitor (%) = 100-C ^ f¾ | fx 100) Determination of antioxidant activity Antioxidant Activity of Cheongho Extract, Octagonal Fennel Extract and Energizer Extract

Using Cayman's Antioxidant Assay Kit, 405 ran inhibited ABTS (2,2'-azino-di- [3-ethylbenzthiazol ine siphonate]) from being oxidized by metmyoglobin to ABTS ⁺ Absorbance was measured and observed.

In other words, the reaction of metomioglobin and hydrogen peroxide generates ferryl myoglobin radicals, which oxidize ABTS to ABTS + with radical cations, which are green at 405 nm. Antioxidants present in the extract of the present invention can suppress the reaction by electron donor radical scavenging to suppress the generation of green ABTS radicals can be represented by measuring the absorbance value. At this time, the amount of antioxidant in the extract of the present invention that inhibits the oxidation of ABTS is indicated by comparing tocope with Trolox analogs.

Into 96 wells, add green leaf extract, octagonal fennel extract or sperm extract 10 ^ and metmyoglobin 10 ≠ (Cayman Chemical Company, MI, USA) and chromogen 150 μί (Cayman Chemical Company, MI, USA), respectively. (Cayman Chemical Company, MI, USA) was added to react. The lid was capped and reacted at room temperature for 5 minutes, and the absorbance was accumulated at 405 ran. All experiment groups were repeated three times to obtain the average absorbance, and the antioxidant activity of the extract of the present invention was the same amount of Troxox standard solution (Trolox standard solution, Cayman Chemical Company, MI, USA) ) Was measured and then compared with the absorbance value of the experimental group to which the sample was placed.励 A receptor affinity test

The affinity of NMDA receptors was determined by measuring the extent of plant extract inhibition of the affinity of [¾] -glycine with NMDA receptors isolated from mouse cerebrum.

Membrane 180 isolated from mouse cerebrum in a 96 well plate and 10 μί, 300 ηΜ [¾] -glycine ((Sigma Aldrich, St. Louis, M0, USA) ) 10 ^ Put in and reacted for 40 minutes on ice. After the reaction, the glass fiber filter (glass fiber filter; 1450-421 Filtermat A) was filtered in a cell harvester (MicroBeta FiltterMate-96 Harvester, USA) and dried in an oven. After drying, the cocktail solution was added and radioactivity was measured using a MicroBeta counter (al lac 1450 MicroBeta counter, Perkin® Elmer, USA) to calculate specific binding. All experimental groups were averaged by performing three replicates, and 1 mM of D-serine was used to calculate non-specific binding. Specific binding of plant extracts and affinity of NMDA receptors were calculated as follows. Specific binding = total binding—non-specific binding

(CPM [sample] — CPM [non-specific texture ¾ ·])

(%) = X 100

(CPM [texture ^■ ]-CPH [non-specific texture ^■ ])

(* CPM ： Counts per minute) Manual Evasion Test and Radial Maze Experiment

Animal behavioral experiments such as passive avoidance and maze experiments were conducted. In the passive avoidance experiment, the control group, scopolamine (3 mg / kg, i.pj-treated group and tacrine (10 mg / kg po) and Chungho extract after scopolamine treatment were administered. After the electric shock was given to the group, the time to enter the dark box was measured 24 hours, and the radial eight-way maze system was checked to confirm the simple memory enhancement effect and the double or more specific learning ability. Acquisition trials were performed to measure the number of errors using (8-arm radial maze).

The experimental results of this study are expressed as mean and standard deviation, and the significance test was performed by Student's t-test using Sigma Plot. Results and Discussion

Measurement of AChE (Acetylcholinesterase) Inhibitory Activity

Acetylcholine is a neurotransmitter that is secreted at the nerve endings and delivers nerve impulses to muscles. After stimulus delivery, acetylcholine is broken down into choline and acetate by AChE.

The hypothesis that cognitive impairment, such as memory loss and learning deterioration caused by senile dementia, is most likely due to acetylcholine neuronal damage in the base of the cerebral base, is currently the most active research and development worldwide and is approved by the FDA. All dementia improvers received were AChE inhibitors. Therefore, in this experiment, we investigated plant resources with AChE inhibitory activity by evaluating enzymatic activity.

The inhibitory activity of the purified AChE from the eel on the clarified ethanol extract, the octagonal fennel ethanol extract, and the energetic ethanol extract was shown in Fig. La-lc, respectively.

Cheongho ethane extract showed lower AChE inhibitory activity compared to 100 fg / u concentration of tacrine, which was used as a positive control. Cheongho (89.44 士 4.05%, 80%, Ethanol extract) at a sample concentration of 1 mg / Showed high levels of enzyme inhibitory activity of more than 80%. It was also observed that all of these samples inhibited enzymatic activity in a concentration-dependent manner at 0.01-10 mg / m ^ concentration (Fig. La). Therefore, combining these results, it was confirmed that the extract of Chungho ethane showed an excellent effect of AChE inhibitory activity.

AChE inhibitory activity of the octagonal ethanol extract is shown in [lb]. Octagonal fennel extracts were tested for acetylcholinesterase inhibitory effects at four concentrations of 0.01 rag / m £, 0.1 mg / l, 1 mg / mi and 10 rag / i, respectively. The inhibitory effect of octagonal fennel was 24.7%, 32.2%, 59.35% and 92.6%, respectively. Inhibitory activity was increased depending on the concentration of the extract. In particular, it showed an inhibitory effect of about 59.35% at a sample concentration of 1 rag /, and the octagonal fennel extract was found to be a relatively good natural product for inhibiting AChE activity.

Compared to tacrine, which used ethanol extract of sperm as a positive control AChE inhibitory activity is shown in [lc]. Energizer extracts were tested for acetylcholinesterase inhibitory effects at four concentrations of 0.01 g / mi, 0.1 mg / m £, 1 mg / mi and 10 rag /, respectively. The inhibitory effect of sperm showed 20.2%, 34.4%, 72.4% and 120% of inhibitory activity, respectively, and the inhibitory activity was increased depending on the concentration of the extract. In particular, it showed an inhibitory effect of about 72.4% at a sample concentration of 1 mg /, sperm extract was also confirmed to be an excellent natural product in inhibiting AChE activity. Determination of Antioxidant Activity

Reactive oxygen species attack lipids, proteins and DNA of cells to produce lipid peroxides and oxidative proteins, promote aging, and cause dementia by causing oxidative damage and destruction of brain cells in brain tissue. It is reported. Therefore, the removal of free radicals is considered to be very important in preventing and treating dementia.

In general, reactive oxygen species occur in series through autooxidation reaction, so the antioxidants in the body terminate this reaction by donating hydrogen atoms to the radicals generated in the autooxidation reaction.

Antioxidants inhibit oxidation by radical scavenging in the electron transport system. In this experiment, we tried to find natural substances that help improve memory by measuring the antioxidant activity of fruit ethanol extract. Antioxidant activity against the plant extracts was measured using the antioxidant assay kit of Cayman's total antioxidant capacity and the experimental results are shown in Figures 2a-2c.

As a result of the measurement of antioxidant activity, green leaf extract was 10-100

Antioxidant activity was increased in a concentration-dependent manner, especially at 10 i / v concentrations of chungho ethane extract (Trolox 0.33 mM or more) showed higher levels of antioxidant activity compared to the positive control trolox ( 2a).

The octagonal fennel extract also contained O.D. Low values were confirmed to show significant antioxidant activity (FIG. 2B).

Energizer extracts also showed lower OD values than trolox 0.225 mM It was found to have excellent antioxidant activity (FIG. 2C).

The measurement results of the antioxidant activity showed that the composition of the present invention can be usefully used as a memory enhancing substance. NMDA Receptor Affinity Test

Recently, research has been focused on controlling dementia by preventing damage to brain cells by various excitatory amino acids such as glutamate and NMDA (N-methyl-D-aspartate).

NMDA receptors act as the most important Ca2 + channels in neurons and induce apoptosis of neurons by promoting the influx of Ca2 + into neurons by various NMDA agonists such as NMDA. Therefore, the antagonists of NMDA receptors with excellent efficacy were measured by measuring the affinity between plant extracts and NMDA receptors. [3H] -glycine, which is known as a selective ligand for NMDA receptors isolated from mouse cerebrum and glycine binding site of these receptors, was used as a sample of chungho ethane extract, octagonal ethanol extract and energetic extract, respectively. We investigated the affinity for NMDA receptors as an indicator of the effects of inhibition of binding. Each plant extract of the present invention was diluted to 0.01, 0.1, 1, and 10 g / mi concentrations to search for affinity for the receptor is shown in Figure 3, the extracts of the present invention are all concentration-dependent to the NMDA receptor Combined (FIGS. 3A-3C). In particular, at high concentrations of 10 mg / n ^, chungho extract (62.21% binding) showed a binding capacity of more than 60% NMDA receptors, octagonal extract inhibited receptor binding at 10 rng / to 82.16%. In addition, the receptor affinity of the sperm extract was 36.1, 40.5, 44.1 and 73.3¾% at 0.01, 0.1, 1 and 10 mg / concentration, respectively, and the receptor binding was inhibited to 44.1% at high concentration of 10 mg /.

Based on these results, the search for affinity for the NMDA receptor glycine binding site of natural plants may be an important indicator that can be usefully used in the research and development of dementia treatments in the future. Confirmation of Memory Improvement Effect of Cheongho Extract Using Passive Evacuation Test Animal behavioral experiments including passive evasion and maze experiment were conducted. In the passive avoidance experiment, the control group remembered the electric shock received 24 hours ago and the time to enter the dark box was significantly increased compared to the training test, but treated with scopolamine (3 mg / kg, iP). One group forgot the electrical stratification due to memory loss and quickly moved to the dark box, indicating that the residence time in the bright box was significantly shorter than the control group. On the other hand, after memory impairment with scopolamine, the time to stay in the bright room was significantly increased in the group administered tacrine (10 mg / kg po) as a positive control. The retention time in the bright box in the group treated with Chungho extract for 5 weeks was significantly increased compared to the scopolamine group. Therefore, it was confirmed that chungho ethanol extract has a significant effect on restoring memory damaged by scopolamine. Confirmation of Memory Improvement Effect of Cheongho Extract Using Radial Maze Experiment

Using the radial maze device, which is a device that can verify the higher memory (spatial energy) effect of the simple memory improvement effect confirmed through the passive evasion experiment, more specifically, the effect of Chungho extract on learning and memory Investigate. In the acquisition trial, which measures the number of errors in radial maze learning, the normal group (1.6) showed lower error counts compared to the negative control (3.25), and both the positive control group, tacrine (1.75) and Chungho, were used as scopolamine. The number of errors was lower than that of the group that suppressed short-term memory.

Memory and cognitive decline are known to be closely related to the cholinergic nervous system. Reportedly, scopolamine, an antagonist of muscarinic receptors at postsynapse, is a neurotransmitter released from presynapse. By interfering with the binding of acetylcholine and the muscarinic receptors to temporarily block information transmission, they impair learning and memory. The results of this experiment seemed to have an effect on minimizing memory and cognitive decline by inhibiting the action of scopolamine that interferes with the binding of acetylcholine and muscarinic receptors. Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims

[Patent Claims]

[Claim 11

A food composition for improving memory or learning ability, comprising one or two or more plant extracts selected from the group consisting of Cheongho (Artemisia apiaceae) extract, octagonal fennel (Illicium veru) extract, and energetic (Lepidi apefalu) extract.

[Claim 2]

For preventing or treating cognitive disorders comprising, as an active ingredient, one or more plant extracts selected from the group consisting of Cheongho (Artemisia apiaceae) extract, octagonal fennel (Illicium verum) extract and energetic Lepidium apefalum extract Pharmaceutical compositions.

[Claim 3]

Cheongho (Artemisia apiaceae) for improving cognitive disorders comprising as an active ingredient one or more plant extracts selected from the group consisting of extracts of spring, Illicium verum and epidiuni apefalum extract Food composition

[Claim 4]

The composition according to any one of claims 1 to 3, wherein the composition inhibits the activity of acetylcholinesterase.

[Claim 5]

The composition according to any one of claims 1 to 3, wherein the composition has antioxidant activity.

[Claim 6]

6. The composition of claim 5, wherein the composition inhibits or eliminates the generation of reactive oxygen species.

[Claim 7]

The method of claim 1, wherein the composition is NMDA.

A composition characterized by acting as an antagonist of the (N-methyl-Daspartate) receptor.

[Claim 8]

4. A composition according to claim 2 or 3, wherein the cognitive impairment is dementia, learning disorder, blindness, forgetfulness, aphasia, execution or delirium.

[Claim 9]

The method of claim 2 or 3, wherein the cognitive dysfunction is AIDS-induced dementia, Vincewanger disease, Lewy body dementia, frontal temporal dementia, mild cognitive impairment, multiple infarct dementia, pick disease, semantic dementia, Alzheimer's dementia or A composition characterized in that it is vascular dementia.

[Claim 10]

Memory comprising the step of administering a composition comprising as an active ingredient one or more plant extracts selected from the group consisting of Cheongho (Artemisia apiaceae) extract, Illicium verm) extract and Lepidium ape f alum) extract Or how to improve learning.

[Claim 11]

Cognition comprising administering a composition comprising as an active ingredient one or more plant extracts selected from the group consisting of Cheongho (Artemisia apiaceae) extract, octagonal fennel (Illicium veruni) extract and energetic Lepidiwn ape fa him) extract How to prevent or treat cognitive disorders.

[Claim 12] The composition of claim 10 or 11, wherein the composition inhibits the activity of acetylcholinesterase.

[Claim 13]

The composition of claim 10 or 11, wherein the composition has antioxidant activity.

[Claim 14]

The composition of claim 13, wherein the composition inhibits or eliminates the generation of reactive oxygen species.

[Claim 15]

12. The composition of claim 10 or 11, wherein said composition acts as an antagonist of an N-methyl- Daspartate (NMDA) receptor.

[Claim 16]

12. The composition of claim 11, wherein the cognitive impairment is dementia, learning disability, blindness, forgetfulness, aphasia, executive or delirium.

[Claim 17]

12. The method of claim 11, wherein the cognitive impairment is AIDS-induced dementia, Vincewan's disease, Lewy body dementia, frontal temporal dementia, mild cognitive impairment, multiple infarct dementia, PEEK disease, semantic dementia, Alzheimer's dementia or vascular dementia Characterized in that the composition.