KR20050075508A - Composition comprising the extract of crude drug complex having neuronal cell-protecting activity for preventing and treating the degenerative brain disease - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of cerebral diseases and a method for producing a complex herbal extract comprising a complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw sulfur extract, golden extract and honey having a neuroprotective activity The complex herbal extract is not only excellent in protecting the nerve cell damage induced by cerebral ischemia, but also harmless to the human body, which is a degenerative brain disease caused by neuronal cell death, namely stroke, stroke, dementia, and Alzheimer's disease. , Parkinson's disease, Huntington's disease, Pick disease and Creutzfeld-Jakob disease can be used as a medicine and dietary supplement for preventing and treating.

Description

Composition comprising the extract of crude drug complex having neuronal cell-protecting activity for preventing and treating the degenerative brain disease}

The present invention relates to a composition for the prevention and treatment of cerebral diseases and a method for producing a complex herbal extract comprising a complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw yellow sulfur extract, golden extract and honey with neuronal cell protective activity will be.

Looking at the proportion of Korea's aging population, announced by the National Statistical Office in October 2003, Korea entered the aging society as the proportion of the population aged 65 or over reached 7.2% in 2000, and this ratio is 14% in 2019. It is expected to enter aging society beyond. As the aging problem becomes a social issue in recent years, the public's interest in the characteristics of the elderly population, the elderly, such as housing, health, culture, leisure, etc. is increasing, and the demand for statistics is increasing. The key to these changes is that chronic degenerative diseases are becoming more of an issue than acute infectious diseases, which have been the main cause of death over the past 50 years due to the aging population. In particular, the death from cerebrovascular disease among chronic degenerative diseases is a very important disease that ranks second in the mortality rate from a single disease.

Cerebrovascular diseases can be classified into two types. One is hemorrhagic brain disease seen in cerebral hemorrhage and the like, and the other is ischemic brain disease caused by occlusion of cerebrovascular vessels. Hemorrhagic brain disease is mainly caused by traffic accidents, and ischemic brain disease is a disease that frequently occurs in elderly people.

In the case of transient cerebral ischemia in the cerebrum, the supply of oxygen and glucose is blocked, resulting in a decrease in ATP and edema in neurons, which eventually leads to extensive brain damage. Neuronal death occurs after a significant period of time after cerebral ischemia, which is called delayed neuronal death. Delayed neuronal death was observed in a transient forebrain ischemic model using Mongolian gerbil, and neuronal cell death was observed in the CA1 region of the hippocampus (hippcampus) 4 days after induction of cerebral ischemia for 5 minutes. (Kirino T, Sano K. Acta Neuropathol., 62 , pp201-208, 1984; Kirino T. Brain Res ., 239 , pp57-69, 1982).

There are two mechanisms of neuronal death caused by cerebral ischemia that most people accept so far. One is the excitatory neuronal death mechanism in which excess glutamate accumulates outside the cell by cerebral ischemia, and this glutamate enters the cell and eventually causes neuronal death due to the accumulation of excess intracellular calcium (Kang TC, et al., J. Neurocytol., 30 , pp945-955, 2001) and two mechanisms of oxidative neuronal death, which are caused by damage to DNA and cytoplasm due to an increase in radicals in vivo due to sudden oxygen supply during ischemia-reperfusion. (Won MH, et al., Brain Res ., 836 , pp 70-78, 1999; Sun AY, Chen Y M. J. Biomed. Sci ., 5 , pp401-414, 1998; Flowers F, Zimmerman JJ. New Horiz. 6 , pp 169-180, 1998).

Based on these mechanisms, many researches have been conducted to search for substances that effectively inhibit neuronal cell death in cerebral ischemia or to reveal the mechanism of the substances. However, there are few substances that effectively inhibit neuronal cell death caused by cerebral ischemia.

Tissue plasminogen activator (Tissue plasminogen activator), the only commercially available drug for treating cerebral ischemia, is a substance that induces rapid supply of oxygen and glucose by melting blood clots that cause cerebral ischemia with thrombolytics. Therefore, because it does not directly protect nerve cells, it is necessary to use it quickly, and because of its characteristic of thrombolytic release, excessive use or frequent use of the thin wall of the blood vessels will eventually lead to hemorrhagic cerebrovascular disease.

MK-801, a calcium channel blocker to effectively inhibit early calcium influx, has been clinically tested, but the drug has been discarded due to its side effects.

On the other hand, in Korea, many natural substances are marketed as health foods that are effective in preventing stroke, but most of them are not scientifically verified, and they are often social problems because they cause health food abuse.

Therefore, there is an urgent need for an effort to objectively verify natural resources that have been used for a long time and to prove their safety, and to develop natural materials having the effect of treating and preventing brain diseases.

Zhaoga is also known as AcantIopanax Senticosus HARMAS, and it is known as Ogapi or Ogalpi in Korea, and Ogapi is a species of arboraceae, but the thorns are called thorn ogapi. In animal experiments, thorny scabies have shown a function and effect to enhance the resistance of living organisms with severe blood circulation disorders and oxygen deficiency symptoms, and have increased resistance to physical (cold, heat and circulation, forced immobilization), chemical and biological adverse effects. In addition to this, it was effective in forming large amounts of antibodies in the body.

Baekbokryeong is a dried core of Poria cocas WOLF. It is also known as Whitesol Pungryeong , and it is grown and cut in pine trees all over the country. It has been used as a tonic since ancient times, and it has the effect of securing the spleen, cutting phlegm, and stabilizing the mind.

Saengjihwang ( Rehmannia glutinosa LIBOSCH.) Is a herb belonging to the family Ginsengaceae, its main component is catalpol, a kind of Iridoid glycosides, and has physiological activities such as diuresis and hypogonadism. In particular, the myopia has a tonic tonic effect and is widely used in various herbal prescriptions and herbal preparations for the purpose of treating anemia, weakness and diabetes.

Golden ( Scutellaria baicalensis ) is a perennial herb of dicotyledon and moss growing in the grass of the mountain and is distributed in Korea, China, Mongolia and eastern Siberia. In oriental medicine, roots are used as antipyretic, diuretic, branch, yedam and anti-inflammatory agents, and they are grown as medicinal plants.

Ginseng is a perennial herb belonging to Ogapiaceae, and saponin-based derivative compounds having various physiological activities have main effects, and ginsenoside content is reported to be an indicator of quality. In oriental medicine prescription, it is used for prescription such as pneumococcal digestive medicine, intestinal fatal medicine, and analgesic pain medication. The pharmacological action is hypoglycemic effect, anticancer effect, serum protein synthesis promotion, antibody production promotion effect, central excitability effect, anti-fatigue effect, etc. have.

The present inventors confirmed that the complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw jihwang extract, golden extract and honey has an activity that can significantly inhibit cerebral ischemic neuronal cell death, and completed the present invention based on this. .

The present invention is a harmless to the human body and prevents degenerative brain disease, including a complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw jihwang extract, golden extract and honey that can prevent nerve cell damage caused by cerebral ischemia as an active ingredient And to provide a therapeutic composition and a method for producing the same.

In order to achieve the above object, the present invention prevents and treats degenerative brain disease, which contains a complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw earth sulfur extract, golden extract and honey as an active ingredient having neuronal protective activity. Provide a pharmaceutical composition.

The degenerative brain disease is characterized by induced neuronal cell death, degenerative brain disease caused by neuronal cell death, stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Pick (Pick) disease and Creutzfeldt- Creutzfeld-Jakob disease may be included.

In addition, the living sulfur extract may be used for living sulfur or juice.

Zhaoga extract, ginseng extract, Baekbokryeong extract, raw jihwang extract and golden extract of the complex herbal extracts are water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, Includes those available in water.

Hereinafter, the present invention will be described in detail.

In general, the complex herbal extract of the present invention, first, a certain amount of zhaoga, ginseng, baekbokyeong, saengjihwang well mixed, and then 1 to 20 times, preferably 4 to 10 times of distilled water for 1 to 24 hours, Preferably, the mixture is refluxed for 2 hours and then filtered to obtain an extract by repeating the process 1 to 5 times, preferably 2 times, and then concentrated under reduced pressure to prepare a concentrate in the form of a concentrate, followed by extracting honey from the extract in the concentrate form. After addition and mixing well, it can be obtained by adding gold in the middle and cooling with a water bath for 48 to 120 hours, preferably about 96 hours.

In addition, the complex herbal extract of the present invention (a) Zhaoga, filtered after refluxing ginseng for about 1 to 20 times, preferably 4 to 10 times distilled water for about 1 to 24 hours, preferably about 2 hours A first step of obtaining the extract by repeating the procedure once to five times, preferably two times, and then concentrating under reduced pressure to prepare a concentrate in the form of a concentrate;

(b) To the extract in the form of the concentrate obtained in the first step was added well mixed Baekbokyeong flour and honey with the juice of raw jihwang, add gold in the middle while cooling for 48 to 120 hours, preferably 96 hours and cooled It may also be obtained from a manufacturing method comprising a manufacturing process consisting of a second step.

Accordingly, the present invention provides a complex herbal extract having a neuronal protective activity obtained from the production method.

Additionally, the complex herbal extracts are suspended in water and then lower acetate, chloroform, acetone, dichloromethane, carbon tetrachloride, such as ethyl acetate in a volume of about 1 to 10 times, preferably about 1 to 5 times the suspension. Nonpolar solvents such as the like may be added to fractionate 1 to 5 times, preferably 2 to 4 times to obtain a nonpolar solvent soluble layer and an aqueous layer. It is also possible to further carry out conventional fractionation processes (Harborne JB, Phytochemical methods: A guide to modern techniques of plant analysis., 3rd Ed. , Pp 6-7, 1998).

The complex herbal extract obtained as described above of the present invention was confirmed to protect the neurons against nerve cell damage caused by focal cerebral ischemia in experimental animals induced by middle cerebral artery occlusion, thereby significantly preventing neuronal cell damage caused by cerebral ischemia.

Accordingly, the present invention provides a complex herbal extract having a neuronal protective activity obtained by the production method, which can be used for the prevention and treatment of degenerative brain diseases caused by neuronal cell death.

In addition, each component of the complex herbal extract is a drug that has been used for a long time as a drug extracts of the present invention extracted therefrom also have no problems such as toxicity and side effects.

Accordingly, the present invention provides a pharmaceutical composition for the prevention and treatment of degenerative brain diseases, including the complex herbal extract with neuronal cell protective activity as an active ingredient.

The pharmaceutical composition for the prevention and treatment of degenerative brain diseases containing the complex herbal extract of the present invention comprises 0.001 to 50% by weight of the complex herbal extract based on the total weight of the composition.

The composition comprising the extract of the complex herbal medicine of the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the combination herbal extract of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

The pharmaceutical composition comprising the complex herbal extract according to the present invention may be prepared in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to a conventional method. It can be formulated and used in the form. Carriers, excipients and diluents that may be included in the composition comprising the combination herbal extract 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, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose, and the like. (sucrose), lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the combination herbal extract of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the preferred effect, the complex herbal extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 100 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The combination herbal extract of the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, the complex herbal extract of the present invention can be used as a main, secondary ingredient and food additive of other foods.

In addition, the present invention provides a health functional food comprising the complex herbal extract and food acceptable food supplements exhibiting a preventive and improving effect of degenerative brain disease. Examples of the food to which the complex herbal extract may be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to food or beverages for the purpose of preventing and improving degenerative brain diseases. At this time, the amount of the combined herbal extract in the food or beverage may be added to 0.01 to 15% by weight of the total food weight, the health beverage composition is based on 100 ml in a ratio of 0.02 to 5 g, preferably 0.3 to 1g Can be added.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the complex herbal extracts as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. have. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the complex herbal extract of the present invention may be used in various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors such as flavoring agents, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the complex herbal extracts of the present invention may contain fruit flesh for the production of natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the complex herbal extract of the invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Experimental Examples.

Example 1 Preparation of Complex Herbal Extract (1)

Zhaoga, Ginseng, Baekbok-ryeong, Saengjihwang and Gold were purchased from the nearby Herbal Medicines and used in the experiments after verification by the Department of Oriental Pharmacology, Graduate School of East-West Medical Science, Kyung Hee University. First, 270 g of Zhaoga and 135 g of ginseng were refluxed for 2 hours with 3 L of distilled water for 2 hours, and then the resultant was filtered twice. 300 g of honey was mixed well with 300 g of saenghwanghwang juice, and then added with golden 6.3 g in the middle while stirring for about 96 hours to obtain 317.2 g of the complex herbal extracts were used as a sample.

Example 2. Preparation of Complex Herbal Extract (2)

Zhaoga, Ginseng, Baekbok-ryeong, Saengjihwang and Gold were purchased from the nearby Herbal Medicines and used in the experiments after verification by the Department of Oriental Pharmacology, Graduate School of East-West Medical Science, Kyung Hee University. First, 270 g of zhao, 135 g of ginseng, 90 g of Baekbokryeong, and 300 g of fresh green sulfur were mixed well, followed by reflux extraction with 6 L of distilled water for 2 hours, followed by filtration twice to obtain an extract. 300 g of honey was added to the concentrated solution (600 ml), and the mixture was mixed well. Then, golden 6.3 g was added thereto while cooling for about 96 hours to obtain 317.2 g of the complex herbal extract, which was used as a sample.

Experimental Example 1. Preparation of experimental animals

Eight weeks-old Sprague dowly male rats of about 280 g as experimental animals were purchased from Samthagosa in Korea, and then adapted to the experimental environment for one week with sufficient feed and water.

Experimental Example 2. Protective effect of neurological extracts of cerebral neurons

Endovascular suture using the method of Zea et al. (Zea LE., Et al., Strok e, 20 , pp84-91, 1989) to measure the neuroprotective effect of the complex herbal extract obtained in Example 1 Intraluminal suture method was used. Closing the middle cerebral artery in rats depletes oxygen and energy sources due to the lack of blood flow in the middle cerebral artery dominant region, resulting in cell death. In particular, the necrosis of the striatum and temporal lobe, the central part of the dominant region, is called ischemic core. And when the phenomenon that the cells themselves die in the area in contact with the dominant area but not the central cerebral artery dominant area is called ischemic penumbra (ischemic penumbra). Necrosis begins 3 hours after the middle cerebral artery is closed, and after 6 hours, it becomes more severe and causes cerebral infarction. At 90 minutes and 24 hours, cerebral infarction is completed and reaches maximum from day 3. When measuring the amount of cerebral infarction (% cerebral infarction), the amount of cerebral infarction (correlated infarct volume, mm ³ ) is measured in consideration of the effects of cerebral edema due to cerebral infarction.

First, at the end of the 25 mm 4-0 nylon suture, about 5-8 mm length was coated with silicon, but the diameter was 0.30 mm and used for the experiment. Rats were subjected to general anesthesia with 5% isoflurane in a mixed gas of 70% N ₂ O and 30% O ₂ , and one side was a syringe for blood collection, and the other was a blood pressure probe. ), And blood pressure was observed, and blood glucose and blood gas were analyzed and observed. The skin in the center of the anterior neck was dissected and the right carotid and external carotid arteries (ECA) were carefully separated from the surrounding tissues and nerves. The upper thyroid artery and the laryngeal artery, which are the branches of the external carotid artery, are cauterized with an electrocatheter. The pterygoid palatal artery, the branch of the internal carotid artery, is cauterized. After inserting about 20 mm was fixed with a thread. The skin incision was resealed and then recovered naturally under anesthesia. Immediately and 120 minutes after the induction, the control group was orally administered to the test group, and the herbal extract of Example 1 was administered orally as a sample. At this time, in order to determine the efficacy according to the dose of the drug, the complex herbal extract of Example 1 was diluted orally administered with tertiary distilled water at a volume of 0.3 ml per 100 g of rat weight, respectively, at 80, 400 and 2000 mg / kg. All experimental groups induced the same number of animals on the same day, and surgery was observed while maintaining the body temperature at 37 ± 0.5 ℃. 120 minutes after the operation, the anesthesia was re-analyzed in the same manner as above to retreat the probe. 48 hours after reperfusion, the cervical spinal bone was removed and brains were excised within 2 minutes to obtain 7 sections 2 mm thick. Subsequently, the tissues were sufficiently immersed in a 16 well plate containing 2% triphenyltetrazolium chloride (TTC) and 37 ° C. After leaving for 30 minutes at 4% paraformaldehyde (paraformaldehide) (fixed parade) to observe the tissue. All surgical procedures were performed under the microscope while maintaining 2% and the lamp was heated to prevent the body temperature from dropping below 37 ℃. Tissues were taken one by one with a digital camera and then transferred to a computer, using the image analysis program Optimas 6.5 (Optimas 6.5, Bioscan) to calculate the cerebral infarction (%) by the following equation (1). At this time, the volume of the corrected cerebral infarction (mm ³ ) was calculated by the following equation (2).

Cerebral infarction rate (%) = (A-B / A) × 100

A: volume of normal left hemisphere (mm ³ ), B: volume of corrected cerebral infarction (mm ³ )

Volume of corrected cerebral infarction (mm ³ )

= (Volume of normal left hemisphere)-(normal site volume of damaged hemisphere)

The results of measuring brain tissue and neuroprotective effects of the combined herbal extracts are shown in FIGS. 1A and 1B. Comparing the two sections of brain tissue in Figure 1a, compared to the control group, the group was administered a combination of the herbal extracts of the area of the dead area where the TTC was not stained, and the area of the area that became black red stained with TTC is less Could know. For reference, when the brain tissue is stained with TTC, the area where the tissue is killed due to the injury is not dyed white, and the tissue in the normal area is black red.

In addition, as shown in FIG. 1B, the cerebral infarction rate (%) was 34.7 ± 1,77% in the water-administered group as a control, while the cerebral infarct rate was 20.5 ± 2.85% when 45 mg / kg of minocycline was used as the positive control. In addition, the doses of the combined herbal extracts 80, 400 and 2000 mg / kg showed brain damage of 34.7 ± 4.19%, 25.6 ± 1.86% and 23.4 ± 2.40%, respectively. In particular, when the multi-drug extract 400 and 2000 mg / kg were administered, they showed 29% and 34% of the neuronal tissue protection, respectively, compared with the control group, and 70 and 81% of the minocycline 45 mg / kg. Showed.

Experimental Example 3. The effect of restoring sensory motor function of complex herbal extract

Brain injury also causes a loss of sensory motor function, so the protective (recovery) effect on sensory motor function ataxia, such as balance, motor function and muscle coordination by the brain damage of the complex herbal extract of Example 1 It was measured through an experiment.

3-1. Rotarod test

A rotarod test was performed to evaluate the effects of the combined herbal extract of Example 1 on sensory motor function 20 hours after cerebral ischemia induction. The rotarod test is one of the most closely tested brain injury models in the brain injury model, especially the stroke model and the spinal nerve injury model.

Rota rod from basile was used for the rota rod test, and the experiment was conducted with an acceleration rotation in which RPM increased to 25 in 2 minutes. The rats that caused strokes as experimental animals were evaluated by measuring the time from the start of rotation on the rod to the drop of the rats after placing the rods in the middle of the rod. Five rats were performed for each rat, and the average of three values except the highest value and the lowest value was used as the average.

Figure 1a shows the results of the rotarod test measured 20 hours after the stroke. Figure 1a shows that the control group exercised for 24.7 ± 6.32 seconds while the combined herbal extract showed 15.1 ± 7.14 seconds, 74.1 ± 12.59 seconds and 63.1 ± 10.08 seconds at 80, 400 and 2000 mg / kg, respectively. The dose of kg showed a significant protective effect against sensorimotor dysfunction compared to the control group (P <0.001).

3-2. Prehensile traction test

The method of Welles (Wallace. JE, et al., J. Gerontol. , 35 , pp364-370, 1980) to evaluate the effect of the combined herbal extract of Example 1 on muscle strength of 21 hours after the induction of cerebral ischemia The prehensile traction test was performed. This is a good way to assess the strength of the rat's foot muscles.

First, the rats were suspended on a 3 cm square rod, and the time until falling was measured. Each time, five rats were used to average the three values and used as data.

As a result, as shown in FIG. 2B, the control group sustained 6.6 ± 0.90 seconds, whereas the combined herbal extract showed 7.2 ± 2.08 seconds, 11.0 ± 1.58 seconds and 7.0 ± 2.31 seconds at 80, 400 and 2000 mg / kg administration, respectively. The dose of mg / kg showed a significant protective effect on motor dysfunction compared to the control group (P <0.05).

3-3. Balance beam test

In order to measure the effect of the compound herbal extract of Example 1 on the balance sensation 22 hours after the induction of cerebral ischemia, a balance beam test was carried out using Purunenen et al. (Puurunen K., et al., Neuropharmacology , 40 , pp 597 -606, 2001) was modified as follows with a slight modification. This test measures the balance of the rat and measures the function of the vestibulomotor. It is one of the good tests to measure the brain function damage after stroke.

First, a wooden rod 3 cm wide and 150 cm long was placed about 50 cm above the floor, and the rats were balanced in the center. After watching the behavior of the rats for 30 seconds, three observers scored them. The evaluation criteria of the score are as follows. 0 points; Unbalanced rod, 1 point; Balanced for 30 seconds, with no movement, 2 points; Balanced for 30 seconds, stretching in the longitudinal direction of the rod, 3 points; Walking on the rod in the longitudinal direction with 2 points on the body, with at least 50% of the steps slipping, 4 points; Sliding from 50 to less than 50%; 5 points; Sliding was about 4 to 1 step, and 6 points | pieces: It carried out by setting it as the state which walked freely without a slipping step at 5 points. Three times per rat, the average value was used as data.

As a result, as shown in Fig. 2c, the control group showed 1.5 ± 0.18 points, whereas the combined herbal extracts showed 1.6 ± 0.25, 2.7 ± 0.19 and 2.08 ± 0.23, respectively, at the 80, 400 and 2000 mg / kg administration At doses of 400 and 2000 mg / kg, there was a significant protective effect on motor dysfunction compared to the control group (P <0.001, P <0.05).

3-4. Foot fault test

In order to measure the effect of the complex herbal extract of Example 1 on motor function at 23 hours after stroke induced by Markgraf CG., Et al., Brain Res. , 575 , pp238-246, 1992) The method of the foot fault test was modified slightly as follows. This test is a good way to measure exercise coordination and precise exercise after a stroke.

First, a 50 cm wide, 150 cm long grid of 4 cm spaced wire is placed about 50 cm above the floor, centered in the rat, and then slipped from the entire step of the forefoot to the annulus step, falling into the grid. The average value was used as data by performing 3 times with the ratio of.

As a result, the control group showed a foot slip of 16.0 ± 2.31%, as shown in Figure 2d, whereas the combined herbal extracts were 20.5 ± 6.25%, 10.7 ± 3.05% and 80, 400 and 2000 mg / kg, respectively. 15.4 ± 2.21% showed protection against motor dysfunction compared to the control at 400 and 2000 mg / kg.

Experimental Example 4. Acute Toxicity

4-1. Oral administration

       ICR-based mice and Sprague-Dawley rats were divided into four groups of 10 rats each, followed by oral administration of the multi-drug extracts of the present invention at doses of 100, 250, 500 and 1000 mg / kg, respectively. No deaths occurred in all four groups and no symptoms were apparent in the control group.

4-2. Intraperitoneal administration

       ICR mice (weight 25 ± 5 g) and Sprague dooli rats were divided into four groups of 10 rats each, and the compound herbal extract of the present invention was intraperitoneally administered at doses of 25, 50, 100 and 200 mg / kg, respectively. No toxicities were observed in all four groups, and no symptoms were apparent in all four groups.

       As a result of the experiment, the herbal extract of the present invention was confirmed that there is almost no acute toxicity.

Hereinafter, an example of the preparation of a pharmaceutical composition comprising the extract of the complex herbal medicine of the present invention will be described, but the present invention is not intended to limit the present invention but only to be described in detail.

Formulation Example 1 Preparation of Powder

Combination Herbal Extract Powder 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Combination Herbal Extract Powder 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Combination Herbal Extract 10 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 Injection

Combination Herbal Extract 10 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, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Combination Herbal Extract Powder 20 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, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Drink

Combination Herbal Extract Powder 100mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

0.25 g of vitamin B ₁

0.3 g of vitamin B ₂

Water quantity

After mixing the above components in accordance with the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare the healthy beverage composition of the invention.

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

As described above, the zhaoga extract, ginseng extract, baekbokyeong extract, raw jihwang extract, golden extract and honey complex herbal extract of the present invention not only has an excellent effect of inhibiting neuronal cell death during cerebral ischemia, but also harmless to human body It can be used for the purpose of preventing and treating degenerative brain diseases caused by necrosis of cells.

Figures 1a and 1b is a measure of the neuroprotective neuronal cell damage protective effect of the extract of the herbal medicine, Figure 1a is a diagram of a brain tissue section of the experimental animal rats were cut by TTC stained brain tissue sections taken with a digital camera, Figure 1b is a comparison of brain tissue damage rate (or cerebral infarction rate) in the control group (water administration), positive control group (minocycline administration), the combination drug extract administration group in rats,

Figures 2a to 2d is a measure of the recovery effect on the sensory motor function after the stroke caused by the herbal extracts, Figure 2a is a diagram showing the rotarod test measured 20 hours after the stroke caused, Figure 2b 21 after the stroke caused FIG. 2C is a diagram illustrating a balance band test performed 22 hours after a stroke and FIG. 2D is a diagram of a foot foul test performed at 23 hours after a stroke.

Claims (7)

A pharmaceutical composition for the prevention and treatment of degenerative brain diseases, comprising a complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw jihwang extract, golden extract and honey as an active ingredient having a neuronal protective activity. The pharmaceutical composition according to claim 1, wherein the extract is an extract available in at least one solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixtures thereof. The pharmaceutical composition of claim 1, wherein the degenerative brain disease is stroke, stroke, dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, Pick's disease, or Creutzfeld-Jakob's disease. A dietary supplement comprising a complex herbal extract consisting of zhaoga extract, ginseng extract, baekbokyeong extract, raw sulfur extract, golden extract and honey and neurologically acceptable food supplements having neuronal protective activity. The health functional food of Claim 4 which is a health drink. Zhao ga, ginseng was refluxed with about 1 to 20 times distilled water for about 1 to 24 hours, and then filtered twice to obtain an extract, concentrated under reduced pressure, and prepared in the form of a concentrate, followed by extract in the form of a concentrate. The method for producing a complex herbal extract comprising a step of mixing the Baekbokyeong powder and honey with fresh saenghwang juice, and then adding gold and cooling in the middle while boiling for 48 to 120 hours. Complex herbal extract obtained by the method of claim 6.