KR101380568B1 - Novel use of extract of Pogongyeong - Google Patents

Abstract

The present invention provides a medicament and / or food use for the treatment, amelioration and / or prevention of either forgetfulness or degenerative brain disease of pogongyoung extract. The present invention has the advantage of being able to effectively treat, ameliorate and / or prevent either forgetfulness or degenerative brain disease.

Description

Novel use of extract of Pogongyeong

The present invention relates to a novel use of pogongyoung extract, and more particularly to a new pharmaceutical or food use of poongong extract.

Forgetfulness with memory-cognitive impairments is of particular interest in modern societies, where the average life expectancy of humans increases.

Degenerative brain diseases in which memory impairment is a problem with forgetfulness include stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, or Huntington's disease.

The dementia generally refers to a condition in which brain function is impaired by various causes and cognitive function is continuously and generally degraded, thereby making it difficult to manage daily and social life. Cognitive function refers to a variety of intellectual skills such as memory, language ability, space-time grasping ability, judgment ability and abstract thinking ability. Each cognitive function is closely related to a specific brain region. The causes of dementia include Alzheimer's disease and Parkinson's disease, degenerative brain diseases, hemorrhagic encephalopathy, metabolic diseases, hepatic encephalopathy, Wilson's disease, infectious neuropathy, acquired immunodeficiency, alcohol and alcohol. There are various etiologies, such as drug addiction and brain trauma, and diseases that can cause structural and functional abnormalities in the central nervous system in some form can cause dementia.

Alzheimer's dementia is the most common degenerative brain disease that causes dementia, the most common cause of dementia in the elderly population, and 50-60% of dementia patients older than 65 years are known as Alzheimer's dementia. The disease develops very slowly and is characterized by a progressive course. In the early stages, they usually showed problems in their memory of recent work, accompanied by abnormalities of language and judgment, and other cognitive functions, and eventually lost all daily life functions. Alzheimer's disease is often accompanied by mental behavioral symptoms such as personality changes, nervousness, depression, delusions, hallucinations, increased aggression, sleep disorders, and neurological disorders such as stiffness and gait abnormalities. Or physical complications such as faeces, incontinence, infections, and pressure sores. Dementia has a longer life discomfort than other serious diseases such as cancer because of its long-term treatment and the nature of its symptoms, and the physical, psychological and economic burden of managing and protecting patients is also very high. Given the rapid growth of the elderly population and the prevalence of dementia, it is clear that dementia is a challenge that requires social and national solutions.

The dementia research, a representative degenerative brain disease so far, is thought to be the main source of cognitive decline mainly due to extensive degeneration and loss of brain cholinergic neurons, which can partially restore the damaged cognitive ability by increasing the activity of the remaining cholinergic nervous system. Most studies have been carried out on drugs. Drugs that have been developed to treat dementia so far include lecithin as an acetylcholine synthetic precursor, RS-86 as a receptor activator, nicotine, and acetylcholinesterase inhibitor (AchE inhibitor) as Aricept from Pfizer / Eisai, Cognex from First Horizon, Exelon from Norvatis, and Reminyl from Janssen. These drugs have the effect of preventing the breakdown of neurotransmitters called acetylcholine, which improves cognitive decline, but the effects are temporary, mild, and seriously toxic. There are many states.

Therefore, there is an urgent need to develop techniques for the treatment, improvement or prevention of forgetfulness and degenerative brain diseases involving memory cognitive impairment.

On the other hand, Pogongyoung (蒲公英) is a herbal medicine that is stored in the Korean herbal medicine (Herbal Medicine) standard collection, it is known that pogongyoung extract is effective in the treatment and prevention of liver disease (see Republic of Korea Patent Publication No. 10-2004-0076106).

Republic of Korea Patent Publication No. 10-2004-0076106, 2004.8.31, claims

The problem to be solved by the present invention is to provide a novel use of the material that is effective in the treatment, improvement or prevention of forgetfulness or degenerative brain disease.

The object of the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

 The inventors have surprisingly found that pogongyoung extract is excellent in improving learning and spatial perception ability of amnesia induced by scopolamine, so that poongong extract is effective in the treatment, improvement and / or prevention of forgetfulness or degenerative brain disease. It was found that the effect was completed to complete the present invention.

The present invention provides a medicament and / or food use for the treatment, amelioration and / or prevention of either forgetfulness or degenerative brain disease of pogongyoung extract.

In another aspect, the present invention provides a pharmaceutical composition for the treatment or prevention of any forgetfulness or degenerative brain disease comprising a pogongyoung extract as an active ingredient.

The pogongyoung extract may be an extract available in at least one solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, and mixtures thereof, and preferably, the poongong extract is an extract available in water or a mixed solvent of water and ethanol. It may be more preferably water or extracts soluble in 5 to 80% ethanol, even more preferably 40 to 80% ethanol.

The degenerative brain disease is not limited thereto, but may be, for example, one or more selected from stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, and Huntington's disease, and preferably may be vascular dementia or Alzheimer's dementia.

The degenerative brain disease may be accompanied by memory cognitive impairment.

The composition may include 0.1 to 50% by weight of the active ingredient based on the total weight of the composition, but is not necessarily limited thereto, and may vary depending on the type and progression of the condition and disease of the user.

The composition may further include appropriate carriers, excipients and / or diluents commonly used in pharmaceutical compositions in addition to the active ingredient.

The composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, or the like, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and / or mineral oil. When formulated, it may be formulated with diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. which are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. Or lactose, gelatin and the like can be formulated. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The preferred dosage of the composition varies depending on the condition and the weight of the patient, the degree of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. However, for the desired effect, it is preferable to administer at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day based on the active ingredient. The administration may be carried out once a day or divided into several doses. Therefore, the dose is not intended to limit the scope of the present invention in any aspect.

The composition can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example by oral and rectal or intravenous methods.

In another aspect, the present invention provides a food composition for improving or preventing any of forgetfulness or degenerative brain disease comprising a pogongyoung extract as an active ingredient.

The food may be in the form of candy, beverage, gum, tea, or the like.

In addition, the food composition may be a health functional food, or a food additive.

The health functional food may be in the form of powder, granules, tablets, capsules or beverages.

Unless otherwise stated, the food composition is equally applicable unless the contradictions mentioned in the pharmaceutical composition of the present invention are contradictory. The term 'improvement' is included in the 'treatment' and means that the condition or symptom is improved.

The food composition may be included in a variety of foods, including beverages. The health functional food refers to a food manufactured using a raw material or ingredient having functional functionality useful for the human body according to the Korean Health Functional Food Act No. 10219 (including processing. By means of nutrient control or physiological effects on the structure and function of the human body to obtain useful effects for health purposes. The food composition may include a conventional food additive, the food additive is a natural chemical such as ketones, glycine, sodium citrate, nicotinic acid, cinnamon acid, navy, licorice extract, crystalline cellulose, high pigment, guar gum And mixed preparations such as additives, sodium L-glutamate preparations, noodle addition alkalis, preservatives, and tar dyes.

Pogongyoung extract may be added to food (including beverages) for the purpose of preventing and / or ameliorating forgetfulness or degenerative brain disease. At this time, the amount of the extract in the food may generally be added in 0.01 to 15% by weight of the total food weight, the beverage composition may be added in a ratio of 0.02 to 10 g, preferably 0.3 to 1 g based on 100 ml. have.

The beverage composition, in addition to containing the extract as an essential ingredient in the indicated ratio, there is no particular limitation on the liquid component and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrin, cyclodextrin and the like Sugar, and sugar alcohols such as xylitol, sorbitol, and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have. The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the method for obtaining the extract used in the use of the present invention or included in the composition will be described in more detail.

For example, the pogongyoung extract may be washed and dried, followed by about 1 to 30 times the weight of the sample, preferably about 5 to 15 times the volume of water (v / w), lower alcohol having 1 to 4 carbon atoms. Or a mixed solvent thereof, preferably water or water and an ethanol mixed solvent, more preferably water or 5 to 80% ethanol, even more preferably 40 to 80% ethanol as the extraction solvent, about 30 to Celsius Conventional extraction methods such as heat extraction, ultrasonic extraction and reflux extraction, preferably ultrasonic extraction, for about 30 minutes to 6 hours, preferably 1 hour to 3 hours at a reaction temperature of 120 degrees, preferably 40 to 80 degrees. After repeated extraction 1 to 10 times, preferably 2 to 7 times, the extract was filtered and concentrated under reduced pressure, and the concentrated extract was lyophilized at about -50 to -30 degrees for about 24 to 72 hours. It can deukhal.

In addition, the pogongyoung extract may be formulated by adding a pharmaceutically or food-acceptable carrier, excipient or diluent, etc. For formulation information, Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA, or See, eg, Food Code.

The present invention can effectively treat, ameliorate and / or prevent either forgetfulness or degenerative brain disease.

1 is a graph showing the results of confirming the memory improvement effect of pogongyoung extract in manual avoidance experiment.
2 to 3 are graphs showing the results of the memory improvement effect of the pogongyoung extract confirmed in Morris water maze experiment.
4 to 5 are graphs showing the results of confirming the memory improvement effect of the pogongyoung extract in the Y maze experiment.
6 is a graph confirming that pogongyoung extract has an effect of improving the memory and cognitive ability by inhibiting the activity of acetylcholinesterase.

Hereinafter, the present invention will be described in more detail with reference to Examples and Preparation Examples, but the following Examples and Formulation Examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

In the present invention, pogongyoung (의미 公 英) is meant to encompass the outposts of Dandelion Taraxacum platycarpum H. Dahlstedt, Taraxacum offficinale Weber, Taraxacum mongolicum Handel-Mazzetti, or Taraxacum coreanum Nakai (Chrysanthemum Compositae). See the Pharmacopoeia Herbal Medicine Specification).

Using a memory decay model (scopolamine treatment model) that characterizes degenerative brain disorders, especially those with memory cognitive impairment, such as forgetfulness to stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, or Huntington's disease, It was confirmed by the following method to have a treatment, improvement and / or prevention effect of forgetfulness to degenerative brain disease.

In the following examples, scopolamine (scopolamin; S1875) and tacrine (tacrine; A3773) are described in Sigma-Aldrich Chemistry Co. The reagents were purchased from and used, and the best reagents commercially available were used. Pogongyoung was purchased from Guilin Pharmaceutical Co., Ltd. (Daegu, Korea).

In addition, the experimental animals were supplied with 6-week-old ICR mice (26-28g) from Daehan Biolink (Chungbuk, Korea) and used for 5 days in a clean cage at Daegu Haany University College of Oriental Medicine. Water and feed were allowed freely, and the temperature (23 ± 2 degrees Celsius), humidity (55 ± 10%) and contrast cycle (12 hours) were automatically controlled.

On the other hand, all experimental results were statistically processed using one-way ANOVA (one way analysis of variance), and the significance test was conducted at p <0.05 level using Student-Newman-Keuls Test when significance was recognized.

<Example 1> In the passive avoidance experiment, confirmed the treatment, improvement and / or preventive effect of forgetfulness to degenerative brain disease of pogongyoung extract

In order to confirm the treatment, improvement and / or prophylactic effect of forgetfulness to degenerative brain disease in passive evacuation experiment of pogongyoung extract (WTP), the experiments were carried out as follows (Kim DH, Jeon SJ, Son KH, Jung JW, Lee S, Yoon BH, Lee JJ, Cho YW, Cheong JH, Ko KH, Ryu JH.The ameliorating effect of oroxylin A on scopolamine-induced memory impairment in mice, Neurobiol Learn Mem, 87, pp. 536-546, 2007).

1-1. Pogongyoung extract preparation

Buy 100 g of dried pogongyoung from Guilin Pharmaceutical Co., Ltd. (Daegu, Korea), dry it completely, grind it, add 10 times (v / w) of distilled water, and use 80 ~ 100 degrees Celsius in reflux cooling system (pyrex, CD1100-300). After extraction, extraction was completed, the extract was filtered with Whatman (No. 1) filter paper, and the filtrate was concentrated under reduced pressure and lyophilized (Eyela, model FDU-2000, Japan) until it became a certain volume. 22.3 g of a water extract (hereinafter referred to as WTP) were obtained (yield: 22.3%).

1-2. Manual Evasion Experiment

Experimental animals prepared at various doses (0, 6.25, 12.5, 25, 50 mg / kg) after dissolving pogongyoung extract (WTP) prepared in 1-1 in 10% Tween 80 (Polyoxyethyl enesorbitan monooleate: Sigma, USA) Administered orally. Positive control group was administered tacrine 10 mg / kg, control group was administered 10% Tween.

Thereafter, the training trial was conducted before the test trial in the following manner.

30 minutes after drug administration, scopolamine dissolved in distilled water was intraperitoneally administered at a dose of 1 mg / kg, and 30 minutes after scopolamine administration, the animals were placed in a brightly lit compartment with a guilotine after 20 seconds of search time. The door was opened to allow access to the dark compartment. Mice that did not enter the dark side within 40 seconds after the guillotine door was opened were excluded from the experiment. After the guillotine door was opened, the time until the rat entered the dark side was measured. Once the rat enters the dark side, the guillotine door closes and a 0.5 mA electric shock flows through the bottom of the grid for three seconds, and the mouse remembers this electric shock.

After this learning test, a test trial was conducted 24 hours later. After 20 seconds of searching time, the time required for the guillotine door to open and four shots to the dark side (latency time) was measured up to 180 seconds. The longer the time taken, the better the learning and memory of passive avoidance.

The results are shown in Fig.

1 is a graph showing the improvement effect according to the dose (6.25, 12.5, 25, and 50 mg / kg) of pogongyoung extract (WTP) in the scopolamine-induced memory loss model in a passive avoidance experiment. In FIG. 1, 10 mice were used in each group, and the retention time was expressed as the mean ± standard deviation.

As shown in the figure, the retention time was significantly reduced when all the experimental groups were administered scopolamine compared to the control group, it can be seen that the memory loss model was produced. In addition, in the dose-dependent experiment of pogongyoung extract (WTP), as shown in Fig. 1, the retention time of the scopolamine-only group was 31.1 ± 5.0 seconds, whereas 119 ± 29.6 ( p <0.05) in the 25 and 50 mg / kg administration groups, respectively. ), 129 ± 15.1 ( p <0.05) seconds.

These results indicate that pogongyoung extract has a dose-dependent effect on memory loss. Thus, not only is it possible to treat or improve degenerative brain diseases involving memory cognitive impairment, such as forgetfulness to stroke, stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, or Huntington's disease associated with memory loss, but also worsen the symptoms The membrane also appears to have a prophylactic effect.

As a result, it can be seen that pogongyoung extract can effectively treat, ameliorate or prevent forgetfulness and / or degenerative brain disease.

Example 2 In Morris Morris Maze Experiment, Confirmation of Treatment, Improvement and / or Prevention of Forgetfulness to Degenerative Brain Disease of Pogongyoung Extract

In order to confirm the treatment, improvement and / or prophylactic effect of forgetfulness to degenerative brain disease in Morris water maze experiment of pogongyoung extract (WTP), experiments were performed as follows (Morris R. Developments of a water-maze procedure for studying spatial learning in the rat.J. Neurosci.Methods.pp. 47-60.1984).

2-1. Pogongyoung extract preparation

A pogongyoung extract was prepared in the same manner as in Example 1-1.

2-2. Morris Underwater Maze Experiment

The pogongyoung extract (WTP) prepared in 2-1 above was dissolved in 10% Tween 80 (Polyoxyethyl enesorbitan monooleate: Sigma, USA) and then orally in experimental animals at various doses (0, 6.25, 12.5, 25, 50 mg / kg). Administered. Positive control group was administered tacrine 10 mg / kg, control group (control) was administered 10% Tween 80. In each of the pogongyoung extract administration group and the positive control group, scopolamine dissolved in distilled water was intraperitoneally administered for 4 days at a dose of 1 mg / kg for 30 minutes after drug administration. (Morris water-maze test) was performed. The aquatic labyrinth device is 90 cm in diameter and 45 cm in height, and the white platform is 6 cm in diameter. The periphery of the underwater maze kept the space cues, such as the video system and the computer system (Ethovision, Noldus, Netherlands) and the device for controlling the water temperature at all times.In the experimental method, the maze was filled with water so that the height of the water was 30 cm. The platform was installed below 1 cm of water height so that the platform could not be seen. When the mouse went to the platform and stayed at the platform for more than 10 seconds, the time taken to find the escape latency was used as the data. At this time, if the average escape latency exceeds 60 seconds, the same value was taken as 60 seconds. On the last 5 days, the platform was removed from the maze and the time the mouse stayed at the platform was measured. Each group used 10 mice.

2 and 3 show the results.

2 to 3 show the improvement of memory of 6.25, 12.5, 25, 50 mg / kg of pogongyoung extract (WTP) in the scopolamine-induced memory decay model in Morris underwater maze experiment, Figure 2 is 4 days Figure 3 is a graph showing the change in latency time (Latency time), Figure 3 is a graph showing the time (Swimming time in target quadrant) the mouse stayed in the position where the platform was on the 5th day.

As shown in Figure 2, as learning progress in the learning test to measure the time required to reach the platform within 60 seconds for 4 days in Morris underwater maze learning, the control (control) is 13.1 ± 2.6 The scopolamine treated group (Scopolamine) rarely found a platform (55.2 ± 1.8 seconds), whereas the OWG-treated group had doses of 25, 50 mg / kg, 20.8 ± 2.2 and 15 ± 1.2, respectively. The time to visit the platform was found to be reduced by seconds ( p <0.05).

In addition, as shown in Figure 3, the scopolamine damaged group (WTP 0 mg / kg, scopolamine 1 mg / kg administration group in the pogongyoung extract (WTP) 25, 50 mg / kg administration group also in the memory test on the fifth day, the last day in the Morris underwater maze learning ) Was significantly increased ( p <0.05).

From these results, it can be seen that pogongyoung extract has an effect on memory loss. Thus, not only is it possible to treat or improve degenerative brain diseases involving memory cognitive impairment, such as forgetfulness to stroke, stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, or Huntington's disease associated with memory loss, but also worsen the symptoms The membrane also appears to have a prophylactic effect.

As a result, it can be seen that pogongyoung extract can effectively treat, ameliorate or prevent forgetfulness and / or degenerative brain disease.

Example 3 Confirmation of the Treatment, Improvement and / or Prevention Effect of Forgetfulness to Degenerative Brain Disease in the Y-maze Test

In order to confirm the effect of treating, ameliorating and / or preventing the forgetfulness to degenerative brain disease of the pogongyoung extract in the Y maze test of the pogongyoung extract (WTP), the experiments were conducted as described below (Sarter M, Bodewitz G, Stephens DN.Attenuation of scopolamineinduced impairment of spontaneous alternation behavior by antagonist but not inverse agonist and betacarboline.Psycopharmacology.pp. 491-495. 1998).

3-1. Pogongyoung extract preparation

A pogongyoung extract was prepared in the same manner as in Example 1-1.

3-2. Y maze test

The pogongyoung extract (WTP) prepared in 3-1. Was dissolved in 10% Tween 80 (Polyoxyethyl enesorbitan monooleate: Sigma, USA) and administered orally at various doses (0, 6.25, 12.5, 25, 50 mg / kg). . A positive control group was administered tacrine 10 mg / kg and a control group was administered 10% Tween 80. Each group of pogongyoung extract and positive control group was intraperitoneally administered scopolamine dissolved in distilled water at a dose of 1 mg / kg 30 minutes after administration of the drug, and then performed a Y-maze test (Y-maze test) It was.

The instrument used for the Y-maze test consists of three branches, each arm is 42 cm long, 3 cm wide, 12 cm high and the angle of contact between the three is 120 degrees. All experimental devices consisted of black polyvinyl plastic. Each branch was set to A, B, and C, and the mouse was carefully placed on one branch, and allowed to move freely for 8 minutes, and then the branch containing the mouse was recorded. Only when the tail is completely entered, it is also recorded when the branch went back. One point (actual alteration) was given to three different branches in turn. Alteration behavior (spontaneous alternation) is defined as entering all three in turn, and is calculated by Equation 1 below. Each group used 10 mice.

Equation  One

% Of change behavior = actual alteration / maximum alterantion × 100 (maximum change: total number of entrances-2)

4 and 5 show the results.

4 to 5 show the improvement effect according to the dose (6.25, 12.5, 25, and 50 mg / kg) of pogongyoung 70% ethanol extract (WTP) in the scopolamine-induced memory loss model in the Y maze experiment 4 is a result graph showing the total entry number by administration group, and FIG. 5 is a result graph showing the spontaneous alternation ratio indicating the alteration behavior by administration group.

As a result, spontaneous alternation was statistically significantly reduced by the administration of scopolamine (1 mg / kg, ip) compared to the control group, and spontaneous alternation was reduced by the administration of 25 and 50 mg / kg polipoglutin extract (WTP). Compared with the administration group (0 mg / kg WTP + Scopolamine 1 mg / kg administration group) it was confirmed that the learning and memory was recovered ( p <0.05).

From these results, it can be seen that pogongyoung extract has an effect on memory loss. Thus, not only is it possible to treat or improve degenerative brain diseases involving memory cognitive impairment, such as forgetfulness to stroke, stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, or Huntington's disease associated with memory loss, but also worsen the symptoms The membrane also appears to have a prophylactic effect.

As a result, it can be seen that pogongyoung extract can effectively treat, ameliorate and / or prevent forgetfulness or degenerative brain disease.

Example 4 Confirmation of Treatment, Improvement and / or Preventive Effect of Forgetfulness to Degenerative Brain Disease by Inhibiting Acetylcholinesterase Activity of Pogongyoung Extract

In order to confirm the effect of treatment, improvement and / or prevention of forgetfulness to degenerative brain disease by inhibiting the acetylcholinesterase activity of the pogongyoung extract (WTP), experiments were conducted as follows (Ellman GL, courtney KD, Andres Jr V and Feather-Stone RM.A new and rapid colormetric determination of acetylcholinesterase activity.Biochem Pharmacol. 7; 88-95.1961).

4-1. Pogongyoung extract preparation

A pogongyoung extract was prepared in the same manner as in Example 1-1.

4-2. Brain tissue acetylcholinesterase (AchE) activity measurement

(1) Reagent

Acetylcholinesterase, acetylthiocholine iodide, 5,5-dithio-bis- (2-nitrobenzoic acid), neostigmine bromide used to measure the activity of acetylcholinesterase bromide) was purchased from Sigma-Aldrich Chemistry Co. and used.

(2) Experimental method

The pogongyoung extract (WTP) prepared in 4-1. And tacrine, a positive control, were dissolved in PBS, and each sample was prepared to have a dose of 0.01, 0.1, 10, 100, 1000 ug / ml, respectively.

Acetylcholinesterase inhibitory activity was measured according to the Ellman method (Ellman et al., 1961). The brains of the mice were extracted and placed in a 10-fold volume of phosphate buffer solution (12.5 mM sodium phosphate buffer pH 7.0, 400 mM NaCl), and the brain tissues were ground using a Teflon glass tube and centrifuged at 1000 xg for 10 minutes at 4 ° C. Supernatants were used as enzymes. 10 μl of the prepared drug and 640 μl of phosphate buffer solution (12.5 mM sodium phosphate buffer pH 7.0, 400 mM NaCl), 25 μl of Ellman's reagent (buffered Ellman's reagent: 10 mM DTNB (5.5'-dithiobis-bis (2-nitrobenzoic acid)) After mixing all 100 μl of enzyme solution, pre-incubate for 30 minutes at room temperature, add 5 μl of 75mM acetylthiocholine iodide and incubate for 10 minutes at room temperature, then add 10 μl of stop reagent Neostigmine bromide (Sigma, USA). After absorbance (OD) was measured at 410nm.

Acetylcholinesterase inhibitory activity was calculated by the following equation.

6 shows the result.

Figure 6 shows the experimental results of the inhibition of acetylcholinesterase activity of the siphon extract (WTP), the x-axis shows the concentration (ug / ml) of the siege extract and tacrine, the y-axis shows the inhibition rate of AchE activity (%) .

As a result, the positive control (tacrine) inhibited acetylcholinesterase activity in a dose-dependent manner, and pogongyoung extract also inhibited acetylcholinesterase activity in a dose-dependent manner, like tacrine. IC ₅₀ value was 1.119 ug / ml.

From these results, pogongyoung extract can be seen to exhibit the effect of improving the memory and cognitive ability by inhibiting the acetylcholinesterase activity like tacrine. Thus, not only is it possible to treat or improve degenerative brain diseases involving memory cognitive impairment, such as forgetfulness to stroke, stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, or Huntington's disease associated with memory loss, but also worsen the symptoms The membrane also appears to have a prophylactic effect.

As a result, it can be seen that pogongyoung extract can effectively treat, ameliorate and / or prevent forgetfulness or degenerative brain disease.

Formulation example  One. Sanje  Produce

WTP 200 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2. Preparation of tablets

WTP 200 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components and tableting according to the conventional tablet manufacturing method to prepare a tablet.

Formulation example  3. Preparation of capsules

WTP 200 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

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

Formulation example  4. Preparation of injections

WTP 200 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ , 12H ₂ O 26 mg

According to the conventional method for preparing an injection, it is prepared in the above-mentioned ingredient content per 2 ampoules.

Formulation example  5. Liquid  Produce

WTP 200 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve it, and lemon flavor is added thereto, then the above ingredients are mixed, adjusted to 100 ml by adding purified water, and then filled into a brown bottle and sterilized to prepare a liquid solution. do.

Formulation example  6. Manufacture of health functional foods

WTP 1000 mg

Vitamin mixture proper amount

Vitamin A Acetate 70 ug

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ug

Vitamin C 10 mg

Biotin 10 ug

Nicotinamide 1.7 mg

Folic acid 50 ug

Calcium Pantothenate 0.5 mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

15 mg potassium monophosphate

Dicalcium Phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients may be mixed according to a general health functional food manufacturing method. Next, the granules may be prepared and used for preparing the nutraceutical composition according to a conventional method.

Formulation example  7. Preparation of health functional food (beverages)

WTP 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with the conventional manufacturing method of health functional food (beverage), and then stirred and heated at 85 degrees Celsius for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2L container, sealed sterilization and refrigerated It is stored and then used to make health functional food (beverage) compositions.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.

Claims (9)

A pharmaceutical composition for treating or preventing forgetfulness, comprising a pogongyoung water extract as an active ingredient. delete delete delete delete Food composition for improving or preventing forgetfulness, including poongyoung water extract as an active ingredient. The food composition of claim 6, wherein the food is in the form of candy, beverage, gum, or tea. The food composition of claim 6, wherein the food composition is a health functional food or a food additive. The food composition of claim 8, wherein the health functional food is in the form of a powder, granule, tablet, capsule or beverage.

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