KR20200038016A - Pharmaceutical composition comprising extract of Sedum kamtschaticum Fisch. for the improvement of memory and preventing or treating neurodegenerative disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for improving cognitive ability and preventing or treating degenerative brain diseases, which contains a Persicaria cochinchinensis extract as an active component. More specifically, the present invention relates to a pharmaceutical composition for improving cognitive ability and preventing or treating degenerative brain diseases, which contains a Persicaria cochinchinensis extract having an excellent inhibitory activity effect on acetylcholinesterase (AchE), a NMDA receptor, and butyrylcholinesterase (BuchE), as an active component.

Description

Pharmaceutical composition comprising extract of Sedum kamtschaticum Fisch. for the improvement of memory and preventing or treating neurodegenerative disease}

The present invention relates to a pharmaceutical composition for preventing or treating degenerative brain disease, including improving the cognitive ability, including the extract of the hairs as an active ingredient, in detail, acetylcholinesterase (AchE), NMDA receptor and butyrylcholinestera (BuchE) relates to a pharmaceutical composition for preventing or treating degenerative brain disease, and improving cognitive ability, which includes an extract having excellent hair inhibitory activity as an active ingredient.

The causes of senile dementia, Alzheimer's disease (AD), which accounts for the largest portion of 71% of all types of dementia, have yet to be identified, but when the brain tissue of Alzheimer's disease patients is observed, the damage to cholinergic nerves It is confirmed that something serious happened. The theory explaining the cause of Alzheimer's disease according to the damage of the cholinergic nerve is called the Cholinergic Hypothesis in Alzheimers disease, and based on this, the activity of acetylcholinesterase that induces a decrease in acetylcholine function is based on this. Many attempts have been made to inhibit it. Drugs currently used as treatments for Alzheimer's disease include tacrine, donepezil, rivastimine and galantamine as inhibitors of acetylcholinesterase. Of these, galantamine is the most recently used with FDA approval. Galantamine prevents the breakdown of acetylcholine and maintains the concentration of acetylcholine at the synapse, thereby improving the cognitive function. However, Galantamine is suspected of its efficacy and safety due to its toxicity and side effects, and these side effects increase public suspicion and fear. Therefore, there is a need to develop safer and more effective natural functional materials.

On the other hand, Persicaria orientalis is a perennial plant attached to the nostalgia family, which lives all over the northern hemisphere and is also called the elderly pole. It grows on a roadside or a vacant land and is 1 to 2 meters tall, making it the largest of its inhabitants in Korea. The entire stem is hairy, thick, and the stem is split much at the top. The leaves are alternate, broad egg-shaped or oval-shaped, 10-20 cm long, 7-15 cm wide and quite large. The tip is pointed, the edges are flat, and the hairs fly back and forth. Flowers bloom from July to September with several branches densely on the branches. The length of the inflorescence is about 5 to 12 centimeters, and it falls down like a rice ear. Flowers are red, without petals, and 8 stamens are longer than calyxes. The fruit is a fruit tree about 3 mm long, dark brown, wrapped in calyx, and ripened.

The present inventors have been reported to have no effect on the treatment of diseases related to dementia, so while studying to develop a dementia prevention or treatment and cognitive improvement agent, the extract of the fluffy acetylcholinesterase (AchE) , By confirming that the NMDA receptor and butyrylcholinesterase (BuchE) inhibitory activity is excellent, it can be usefully used as an active ingredient of the pharmaceutical composition for improving cognitive ability and preventing or treating degenerative brain disease. By revealing, the present invention was completed.

The present invention is an acetylcholinesterase (AchE), NMDA receptor and butyrylcholinesterase (BuchE) inhibitory activity is excellent cognitive ability to improve the cognitive ability to contain the extract as an active ingredient, and for preventing or treating degenerative brain disease pharmacy It is an object to provide a composition and a method for manufacturing the same.

In addition, the present invention has another object to provide a health functional food for improving cognitive ability, and preventing or treating degenerative brain disease, which includes the pharmaceutical composition according to the present invention as an active ingredient.

The present invention provides a pharmaceutical composition for improving or preventing cognitive ability, and preventing or treating degenerative brain disease, including the hairy extract as an active ingredient.

According to an embodiment of the present invention, the hairy extract may be extracted from at least one selected from flowers, branches, stems, leaves and roots of hairy leaves.

In addition, according to an embodiment of the present invention, the hairy extract is extracted from the flower of hairy leaves, and may be included in a concentration of 10 to 70 μg / ml in the pharmaceutical composition.

In addition, according to an embodiment of the present invention, the hairy extract is extracted from the root of hairy hairy, it may be included in a concentration of 10 ~ 70 μg / ml in the pharmaceutical composition.

In addition, according to an embodiment of the present invention, the hairy extract is at least any one selected from acetylcholinesterase, NMDA receptor (N-methyl-D-aspartate receptor), and butyrylcholinesterase One can exhibit inhibitory activity.

In addition, according to an embodiment of the present invention, the hairy extract is extracted from the flower of hairy hair, exhibits inhibitory activity on the NMDA receptor (N-methyl-D-aspartate receptor), or the hairy hair extract is extracted from the hairy hairy root, It can exhibit inhibitory activity on acetylcholinesterase and butyrylcholinesterase.

In addition, according to an embodiment of the present invention, the degenerative brain disease is selected from Alzheimer's disease (Alzheimer's disease), Pick disease (Pick disease), Parkinson's disease (Parkinson's disease), Lou Gehrig's disease (amyotrophic lateral sclerosis) and Huntington's disease (Huntington's disease) It can be either.

In addition, the present invention provides a method for improving the cognitive ability, and preparing a pharmaceutical composition for the prevention or treatment of degenerative brain disease, including the step of obtaining the extract by extracting it at a predetermined temperature and pressure condition after mixing it with a solvent. .

According to an embodiment of the present invention, the hairy hairy part of the hairy, the solvent is at least one selected from lower alcohols of C1 to C4, ethyl acetate, acetone, water and hexane, and the hairy extract is 25 to 100 Can be extracted at ℃.

In addition, according to an embodiment of the present invention, the root portion of the fluffy fluffy coat, the solvent is at least one selected from lower alcohols of C1 to C4, ethyl acetate, acetone, water and hexane, and the fluffy extract is 25 ~ It can be extracted at a temperature of 100 ℃ and a pressure of 145 ~ 2000 psi.

In addition, the present invention provides a health functional food for preventing or treating degenerative brain disease, and improving cognitive ability, which includes the hairy extract as an active ingredient.

The pharmaceutical composition comprising the hairy extract according to the present invention as an active ingredient prevents or minimizes the death of brain cells by inhibiting the activity of acetylcholinesterase, NMDA receptor and butyrylcholinesterase (BuchE), and brain By increasing the concentrations of acetylcholine and butyrylcholine secreted from, it can treat or prevent degeneration of cholinergic neurological function, and thus can be widely used as a drug or health functional food for preventing or treating dementia and improving cognitive function.

Figure 1a is a graph showing the results of measuring the cytotoxicity of the hairy extract prepared in Example 1.
Figure 1b is a graph showing the results of measuring the cytotoxicity of the hairy extract prepared in Example 5.
1C is a graph showing the results of cytotoxicity measurement for the hairy extract prepared in Example 9.

In order to solve the above-mentioned problems, the present invention prevents, treats, and recognizes dementia including an active ingredient containing an extract that has excellent activity inhibiting the activity of acetylcholinesterase, NMDA receptor, and butyrylcholinesterase (BuchE) as an active ingredient. It is intended to provide a pharmaceutical composition for improving ability.

The term “composition” refers to a mixture of the hairy extract of the present invention with other chemical components such as diluents or carriers.

The term “carrier” is defined as a compound that facilitates the addition of a compound into a cell or tissue. For example, dimethyl sulfoxide (DMSO) is a commonly used carrier that facilitates the introduction of many organic compounds into a cell or tissue of an organism.

The term “diluent” is defined as a compound that dilutes in water that will dissolve the compound, as well as stabilize the biologically active form of the target compound. Salts dissolved in buffer solutions are used in the art as diluents. A commonly used buffer solution is phosphate buffered saline, because it mimics the salt state of human solutions. Because buffer salts can control the pH of a solution at low concentrations, buffer diluents rarely modify the biological activity of the compound.

All technical terms used in the present invention are used in the meaning as generally understood by those skilled in the art in the relevant field of the present invention, unless otherwise defined. In addition, although a preferred method or sample is described herein, similar or equivalent ones are included in the scope of the present invention.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art to which the present invention pertains may easily practice. The present invention can be implemented in many different forms and is not limited to the embodiments described herein.

The present invention provides a pharmaceutical composition for the prevention or treatment of dementia and improvement of cognitive ability, which includes the hairy extract as an active ingredient.

Even substances previously known to have antioxidant efficacy do not have the effect of inhibiting the generation of reactive oxygen species in neurons, and thus do not have the effect of suppressing the death of neurons due to oxidative stress. It has excellent inhibitory activity against acetylcholinesterase and butyrylcholinesterase, increasing the concentration of acetylcholine and butyrylcholine secreted by the brain, causing degenerative brain disease caused by degeneration of cholinergic neurological function It can be prevented or treated, can exhibit a cognitive improvement effect, and by inhibiting the activity of the NMDA receptor to minimize or prevent the death of brain cells, it can exhibit the effect of improving cognition and preventing or treating degenerative brain disease.

The hairy extract can be extracted using any one of the branches, stems, flowers, fruits, or roots of hairy leaves, but among the above-mentioned parts, the extract extracted from the hairy leaves is acetylcholinesterase ) And butyrylcholinesterase, as well as the inhibitory effect of NMDA receptor activity.

The hairy extract extracted from the flowery hairy hair extract can be included in a concentration of 10 to 70 μg / ml in the pharmaceutical composition, and if the hairy hair extract is included in a concentration of less than 10 μg / ml, the inhibitory effect of NMDA receptor activity is It may not be expressed at the desired level, and when it is included in a concentration exceeding 70 μg / ml, the cytotoxicity of the extract may be reduced.

More preferably, the hairy extract extracted from the flowery part of the hairy hair can be included in a concentration of 40 to 60 μg / ml in the pharmaceutical composition, in this case acetylcholinesterase, butyrylcholinesterase (butyrylcholinesterase) and NMDA receptors may have better inhibitory effects.

When the extract is extracted using the branch or root portion of the furrow, more preferably the root portion, the acetylcholinesterase and butyrylcholinesterase (BuchE) activity inhibitory effect may be more excellent.

The hairy extract extracted from the root of the hairy skin can be included in a concentration of 10 to 70 μg / ml in the pharmaceutical composition, and if the hairy skin extract is included in a concentration of less than 10 μg / ml, acetylcholinesterase And butyrylcholinesterase (BuchE) activity inhibitory effect may not be expressed at the desired level, and when it is included in a concentration exceeding 70 μg / ml, the cytotoxicity of the extract can be increased.

More preferably, the hairy extract extracted from the root part of the hairy skin can be included in a concentration of 40 to 60 μg / ml in the pharmaceutical composition, and accordingly, acetylcholinesterase and butyrylcholinesterase (BuchE) The activity inhibitory effect of may be more excellent.

The degenerative brain disease may be any one selected from Alzheimer disease, Pick disease, Parkinson's disease, amyotrophic lateral sclerosis and Huntington's disease.

Next, a method of improving the cognitive ability and preventing or treating degenerative brain disease according to the present invention will be described.

The method for improving the cognitive ability of the present invention and for preparing a pharmaceutical composition for preventing or treating degenerative brain disease includes a step of mixing with a fluffy solvent and extracting it at a predetermined temperature and pressure condition to obtain a fluffy extract.

The shedding may be used as a sample after washing, drying and pulverizing before extraction, and the method for preparing the shedding in powder form may use a pulverization process of herbs commonly used in the art.

When using the flower part of the fur, the solvent may be at least one selected from lower alcohols of C1 to C4, ethyl acetate, acetone, water and hexane, preferably methanol, and the fur extract is extracted at 25 to 100 ° C. Can be.

When the extract temperature is less than 25 ° C when extracting the extract, the content of the active ingredient in the obtained extract of the extract is low, so the acetylcholinesterase (AchE), NMDA receptor, and butyrylcholinesterase (BuchE) inhibitory activity effects are intended. May not be expressed at a level, and when it exceeds 100 ℃, the active ingredient in the obtained hairy extract is destroyed and acetylcholinesterase (AchE), NMDA receptor and butyrylcholinesterase (BuchE) inhibitory activity effect It may be difficult to achieve the object of the present invention, such as not being expressed at the desired level.

Next, when using the root portion of the hairpin, the solvent may be at least one selected from lower alcohols of C1 to C4, ethyl acetate, acetone, water and hexane, preferably ethanol, and the hairy extract is 25 to 100 ℃ and 145 ~ 2000 psi, more preferably 700 ~ 2000psi, even more preferably can be extracted at a pressure of 1500 ~ 2000psi.

When the extract temperature is less than 25 ° C when extracting the extract, the content of the active ingredient in the obtained extract of the extract is low, so the acetylcholinesterase (AchE), NMDA receptor, and butyrylcholinesterase (BuchE) inhibitory activity effects are intended. It may not be expressed at the level, and if it exceeds 100 ℃, there may be a problem such as the content of the active ingredient in the extract is not increased or the active ingredient in the extract is destroyed even if the temperature is increased.

In addition, when the extraction pressure is less than 145psi when extracting the extract, the content of the active ingredient in the obtained extract of the extract is low, so that the acetylcholinesterase (AchE), NMDA receptor, and butyrylcholinesterase (BuchE) inhibitory activity effects are intended. It may not be expressed at the level, and if it exceeds 2000psi, the content of the active ingredient in the extract of the hairs can be increased even if the pressure is increased. When the extraction pressure is 700 ~ 2000psi when extracting the extract, the content of the active ingredient in the extracted extract is further increased, and the effect of preventing or treating degenerative brain disease can be further improved, and the pressure of 1500 ~ 2000psi In one case, the above-described effect can be further improved.

The hairy extract can be obtained in powder form by removing the solvent after the hairy extract is extracted with the solvent, and the method of removing the solvent may use a solvent removal method commonly used in the art, for example, vacuum filtration. However, it is not limited thereto.

Meanwhile, the pharmaceutical composition for improving cognitive ability and preventing or treating degenerative brain disease according to the present invention is an oral dosage form such as powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, etc. It can be used in the form of external preparations, suppositories and sterile injectable solutions.

In addition, the pharmaceutical composition is formulated in a unit dose form by formulating using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily carried out by a person skilled in the art to which the present invention pertains. Or it can be manufactured by incorporating into a multi-dose container.

The pharmaceutical composition for improving the cognitive ability of the present invention and for preventing or treating degenerative brain disease may be included in a drug and used, and the pharmaceutically acceptable carrier included in the drug is commonly used in preparation, lactose , Dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxy Benzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, and the like, but is not limited thereto.

In addition, when the pharmaceutical composition for improving the cognitive ability of the present invention and for preventing or treating degenerative brain disease is used in a pharmaceutical, in addition to the above components, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspensions, preservatives, etc. are additionally added. It can contain. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The drug may be administered orally or parenterally, and for parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, intrapulmonary administration, and rectal administration, etc. You can. When administered orally, proteins or peptides are digested, so oral compositions must be formulated to coat the active agent or to protect it from degradation in the stomach. In addition, the active agent can be administered by any device capable of moving to the target cell.

Suitable dosages of such pharmaceuticals vary by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity, and are usually skilled The doctor can easily determine and prescribe a dose effective for the desired treatment or prevention.

The medicine may be administered as an individual prophylactic or therapeutic agent, or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents.

In addition, the present invention provides a health functional food for improving or improving cognitive ability, including depilatory extract as an active ingredient, and for preventing or treating degenerative brain disease.

The type of the health functional food composition is not particularly limited as long as it is usually manufactured and / or sold. For example, dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes It can be used in the form of pills, powders, granules, needles, tablets, capsules or beverages, and includes all health functional foods in the usual sense.

The healthy beverage composition is not particularly limited to the liquid component except that it contains the hairy extract according to the present invention, and may contain various flavoring agents or natural carbohydrates, etc., as additional components, like a conventional beverage.

Typically, the amount of hairy extract contained in the health functional food may be included in 0.1 to 50% by weight of the total food weight, preferably 1 to 40% by weight. In addition, in the case of long-term intake for the purpose of health and hygiene or for health control, it may be below the above range, and since there is no problem in terms of safety, the active ingredient may also be used in an amount above the above range.

Hereinafter, an example of the preparation of an injection containing the pharmaceutical composition for improving cognitive ability of the present invention and preventing or treating degenerative brain disease as an active ingredient will be described, but the present invention is not intended to be limited thereto, but only to be specifically described. .

<Production Example 1>

Hair extract 10 mg

Mannitol 180 mg

Injectable sterile distilled water 2974 mg

Na ₂ HPO ₄ , 12H ₂ O 26mg

Hereinafter, an example of manufacturing a health functional food for improving cognitive ability of the present invention and preventing or treating degenerative brain disease will be described, but the present invention is not intended to be limited thereto, but only to be specifically described.

<Production Example 2>

200mg of fluffy extract

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B 1 0.13 mg

Vitamin B 2 0.15 mg

Vitamin B 6 0.5mg

Vitamin B 12 0.2 μg

Vitamin C 10 mg

Biotin10 ㎍

Nicotinic acid amide 1.7 mg

50 ㎍ folic acid

Calcium Pantothenate 0.5 mg

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

The present invention will be described in more detail through the following examples. However, the following examples are merely examples for easily explaining the contents and scope of the technical spirit of the present invention, and thus the technical scope of the present invention is not limited or changed. In addition, various modifications and changes are possible within the scope of the technical spirit of the present invention based on these examples can be easily determined by those skilled in the art.

(Example 1)

After mixing the root part of the hair with a mixture of ethanol, it was extracted for 14 minutes at 50 ° C and 1,700 psi pressure using an accelerated solvent extraction device. The solvent was removed at 50 ° C. using a reduced pressure concentrator to obtain an extract. After dissolving the obtained hairy extract in 0.02% DMSO aqueous solution, it was diluted with 100 mM of sodium phosphate buffer to prepare a pharmaceutical composition containing the hairy extract as a concentration of 25 μg / ml.

(Example 2)

A pharmaceutical composition was prepared in the same manner as in Example 1, but containing the extract with a concentration of 50 μg / ml.

(Example 3)

A pharmaceutical composition was prepared in the same manner as in Example 1, but containing the extract with a concentration of 100 μg / ml.

(Example 4)

A pharmaceutical composition was prepared in the same manner as in Example 1, but containing the extract with a concentration of 250 μg / ml.

(Example 5)

The same procedure as in Example 1 was carried out, but the branch portion was used instead of the root portion of the furrow, and the obtained furrow extract was dissolved in 0.02% DMSO aqueous solution and diluted with 100 mM sodium phosphate buffer to obtain the furrow extract. A pharmaceutical composition was prepared containing 25 μg / ml.

(Example 6)

A pharmaceutical composition was prepared in the same manner as in Example 5, but containing the extract with a concentration of 50 μg / ml.

(Example 7)

A pharmaceutical composition was prepared in the same manner as in Example 5, but containing a hairy extract at a concentration of 100 μg / ml.

(Example 8)

A pharmaceutical composition was prepared in the same manner as in Example 5, but containing the extract with a concentration of 250 μg / ml.

(Example 9)

After mixing the flower parts of the hairs with methanol, the mixture was extracted at 74 ° C and atmospheric pressure for 4 hours. After extraction, the solvent was removed at 50 ° C. using a vacuum concentrator to obtain an extract. The obtained hairy extract was dissolved in an aqueous solution of 0.02% DMSO, and then diluted with 100 mM sodium phosphate buffer to prepare a pharmaceutical composition containing the hairy extract as a concentration of 25 μg / ml.

(Example 10)

A pharmaceutical composition was prepared in the same manner as in Example 9, but containing the extract with a concentration of 50 μg / ml.

(Example 11)

A pharmaceutical composition was prepared in the same manner as in Example 9, but containing the extract with a concentration of 100 μg / ml.

(Example 12)

A pharmaceutical composition was prepared in the same manner as in Example 9, but containing the extract with a concentration of 250 μg / ml.

(Comparative Example 1)

The commercial dementia treatment product, tacrine, was diluted with 100 mM of sodium phosphate buffer to prepare a pharmaceutical composition containing the tacrine at a concentration of 0.1 μg / ml.

(Experimental Example 1) Cytotoxicity measurement

Cytotoxicity was measured by dispensing the BV-2 cell line into a 48 well plate at a ratio of 2 × 10 ⁵ cells / ml, followed by culturing for 24 hours, processing the extract sample after shaking, incubating for 24 hours, and adding 0.6 mg / ml MTT reaction solution. After 1 hour after treatment, DMSO was added to dissolve formazan to measure absorbance at a wavelength of 540 nm using a microplate reader. The cytotoxicity measurement results are shown in FIGS. 1A to 1C.

1A to 1C show the results of cytotoxicity measurement for samples containing the hairy extracts prepared in Examples 1, 5, and 9 at concentrations of 12.5, 25, 50, 100, and 200 μg / ml, respectively. It is done.

Referring to Figures 1a to 1c, it can be seen that the cell proliferation rate is reduced at a concentration of 100 μg / ml or more in the roots, branches, and flower parts of the fur.

(Experimental Example 2)

In order to improve the cognitive ability of the hairy extract prepared in Examples and to evaluate the effect of preventing or treating degenerative brain diseases, the inhibitory activity of acetylcholinesterase (AchE), NMDA receptor and butyrylcholinesterase (BuchE) It was evaluated as follows.

1) Evaluation of acetylcholinesterase (AchE) inhibitory activity

Mixture 1 was prepared by diluting the hairpin extract prepared in Example and the talc of Comparative Example in 100 mM sodium phosphate buffer at the concentrations shown in Table 1 below.

Mixture 2 was prepared by adding 10 mM DTNB (5,5'-dithio-bis- [2-nitrobenzoic acid]) and 15 mM sodium bicarbonate to 100 mM sodium phosphate buffer.

Mixture 3 was prepared by mixing mixture 1 (1.5 ml), mixture 2 (100 ml) and 100 mM sodium phosphate buffer (2.6 ml).

To the mixture 3, 75 mM acetylthiocholine iodide (20 ml) was added and left for 10 minutes. Next, 0.3 U / ml of acetylcholinesterase (50 μl) was added and reacted for 30 minutes, and absorbance was measured at 410 nm at 1 minute intervals.

The acetylcholinesterase inhibition rate was calculated by reflecting the absorbance measurement results in the following Equation 1.

<Calculation formula 1>

In the above formula 1, the absorbance of the experimental group is a measurement of absorbance at 410 nm when the extract of Example is used or the talc of comparative example, and the absorbance of the control is the absorbance measurement value of 100 mM sodium phosphate buffer. it means.

2) Evaluation of butyrylcholinesterase (BuchE) inhibitory activity

Mixture 1 was prepared by diluting the hairpin extract prepared in Example and the talc of Comparative Example in 100 mM sodium phosphate buffer at the concentrations shown in Table 1 below.

Mixture 2 was prepared by adding 10 mM DTNB (5,5'-dithio-bis- [2-nitrobenzoic acid]) and 15 mM sodium bicarbonate to 100 mM sodium phosphate buffer.

Mixture 3 was prepared by mixing mixture 1 (1.5 ml), mixture 2 (100 ml) and 100 mM sodium phosphate buffer (2.6 ml).

To the mixture 3, 75 mM butyrylthiocholine iodide (20 ml) was added and left for 10 minutes. Next, 0.5 U / ml butyrylcholinesterase (50 μl) was added and reacted for 30 minutes, and absorbance was measured at 410 nm at 1 minute intervals.

The absorbance measurement result was reflected in the following calculation formula 2 to calculate the butyrylcholinesterase inhibition rate.

<Calculation formula 2>

In the above formula 1, the absorbance of the experimental group is a measurement of absorbance at 410 nm when the extract of Example is used or the talc of comparative example, and the absorbance of the control is the absorbance measurement value of 100 mM sodium phosphate buffer. it means.

division sample part extraction
Condition* Extraction condition density
(μg / ml) Acetylcholinesterase inhibition rate
(%) Butyrylcholinesterase inhibition rate
(%) menstruum Temperature
(℃) pressure
(psi) Example 1 Fluffy Root 26 ethanol 50 1700 25 18.2 35.7 Example 2 Fluffy Root 26 ethanol 50 1700 50 25.5 57.4 Example 3 Fluffy Root 26 ethanol 50 1700 100 46.3 74.8 Example 4 Fluffy Root 26 ethanol 50 1700 250 66.3 79.0 Example 5 Fluffy Branch 26 ethanol 50 1700 25 7.0 22.5 Example 6 Fluffy Branch 26 ethanol 50 1700 50 21.1 25.4 Example 7 Fluffy Branch 26 ethanol 50 1700 100 31.2 48.6 Example 8 Fluffy Branch 26 ethanol 50 1700 250 58.4 71.1 Example 9 Fluffy Flower One Methanol 74 14.696 25 7.7 9.4 Example 10 Fluffy Flower One Methanol 74 14.696 50 11.8 19.7 Example 11 Fluffy Flower One Methanol 74 14.696 100 18.4 50.2 Example 12 Fluffy Flower One Methanol 74 14.696 250 43.9 69.4 Comparative Example 1 Tacrine - - - - - 0.1 76.8 -

Referring to Table 1, on the basis of 100 μg / ml of the hairy extract, the hairy extract extracted from the root part of the hairy extract has a higher inhibition rate of acetylcholinesterase and butyrylcholinesterase than the hairy extract extracted from other parts. It can be confirmed that it is excellent.

3) NMDA receptor inhibitory activity evaluation

Conditions containing or not extracting the cortical homogenate corresponding to 1 mg protein at 4 ℃ with 5 nM of [ ³ H] CGP39653, 5 mM of Tris-HCl (pH7.7) and 10 mM of EDTA-Tris Incubated for 60 minutes.

After incubation, the sample was quickly filtered under vacuum with glass fiber filters (GF / B, Whatman) and washed several times with ice-cold incubation buffer using a 48-sample cell harvester (Brandel).

After the filtrate was dried, radioactivity was calculated in a scintillation counter (Scintillation counter, LS series, Beckman) using a fluorescence solution (scintillation cocktail, Formula 989, Packard), and the calculated results were calculated for control radioligand specific binding. Inhibition rate (%). CGS 19755 was used as a standard reference compound, and the standard comparison compound was used for each experiment at various concentrations to obtain a calibration curve for obtaining IC ₅₀ .

division sample part extraction
Condition* density
(μg / ml) Extraction condition NMDA receptor inhibition rate
(%) menstruum Temperature
(℃) pressure
(psi) Example 10 Fluffy Flower One 50 Methanol 25 1700 50.9 Example 6 Fluffy Branch 26 50 ethanol 50 1700 7.9

Referring to Table 2, it can be seen that, on the basis of 50 μg / ml of the hairy extract, the hairy extract extracted from the middle part of the hairy extract has superior NMDA receptor inhibition rate than the hairy extract extracted from the eggplant.

Claims (11)

A pharmaceutical composition for improving cognitive ability, which includes the extract of the hair as an active ingredient, and for preventing or treating degenerative brain disease.
According to claim 1,
The yeokkyeokkyeo extract is yeokkyeokkyeo flowers, branches, stems, leaves and roots, improved cognitive ability, and a pharmaceutical composition for preventing or treating degenerative brain disease, characterized in that extracted from at least any one site.
According to claim 2,
The hairy extract is extracted from the flower of hairy leaves, improving the cognitive ability included in a concentration of 10 ~ 70 μg / ml in the pharmaceutical composition, and a pharmaceutical composition for preventing or treating degenerative brain disease.
According to claim 2,
The hairy extract is extracted from the root of hairy leaves, improving the cognitive ability included in a concentration of 10 ~ 70 μg / ml in the pharmaceutical composition, and a pharmaceutical composition for preventing or treating degenerative brain disease.
According to claim 1,
The hairy extract is acetylcholinesterase (acetylcholinesterase), NMDA receptor (N-methyl-D-aspartate receptor), and butyrylcholinesterase (butyrylcholinesterase) selected from at least one selected from among the improved cognitive ability to exhibit inhibitory activity, And a pharmaceutical composition for preventing or treating degenerative brain disease.
According to claim 1,
The hairy extract is extracted from the hairy flower, and exhibits an inhibitory activity to the N-methyl-D-aspartate receptor (NMDA) or
The hairy extract is extracted from the roots of hairy leaves, improving cognitive ability showing inhibitory activity on acetylcholinesterase and butyrylcholinesterase, and a pharmaceutical composition for preventing or treating degenerative brain disease.
According to claim 1,
The degenerative brain disease is Alzheimer's disease (Alzheimer disease), Pick disease (Pick disease), Parkinson's disease (Parkinson's disease), Lou Gehrig's disease (amyotrophic lateral sclerosis) and Huntington's disease (Huntington's disease) selected from among cognitive improvement, and degenerative brain Pharmaceutical composition for preventing or treating diseases.
A method for improving the cognitive ability, and preparing a pharmaceutical composition for preventing or treating degenerative brain disease, comprising the step of obtaining a hairy extract by mixing it with a solvent and then extracting it under a predetermined temperature and pressure condition.
The method of claim 8,
The furrow is the flower part of the furrow, and the solvent is at least one selected from lower alcohols of C1 to C4, ethyl acetate, acetone, water, and hexane, and the furrow extract is improved in cognitive ability extracted at 25 to 100 ° C, and Method for preparing pharmaceutical composition for preventing or treating degenerative brain disease.
The method of claim 8,
The furrow is the root portion of the furrow, and the solvent is at least one selected from lower alcohols of C1 to C4, ethyl acetate, acetone, water and hexane, and the furrow extract has a temperature of 25 to 100 ° C and a temperature of 145 to 2000 psi. A method of manufacturing a pharmaceutical composition for improving cognitive ability extracted from pressure and preventing or treating degenerative brain disease.
A health functional food for improving cognitive ability, including depilatory extract as an active ingredient, and for preventing or treating degenerative brain disease.

Images