The present invention relates to a composition comprising an extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI, prevention and treatment of stroke and neurodegenerative diseases such apoplexy, Alzheimer's disease (AD), Parkinson's disease (PD), Pick's disease, Creutzfeldt-Jakob disease and senile dementia.

Description of the Invention

TECHNICAL FIELD

The present invention relates to an extract of crude drug complex having neuroprotective activity for preventing and treating stroke and neurodegenerative diseases.

BACKGROUND ART

In the twentieth century, as the average life span of human has been increasing with the rapid development of life science and medicine, new social problems including increased population ratio of older people are coming to the front, especially, the geriatric neuronal diseases such as stroke, Alzheimer's disease (AD), Parkinson's disease (PD) etc., which are fatal functional disorder of neuronal system, have been increased.

The growth, differentiation and death of neuronal cell in neuronal system are important control processes in general development, the establishment of tissue specific function and the maintenance of homeostasis respectively.

Cerebrovascular disease is classified into two types, hemorrhagic brain disease and ischemic brain disease: the hemorrhagic brain disease such as cerebral hemorrhage occurs mainly by bleeding of cerebral vessels and the ischemic brain disease frequently occurring in older people is caused by the occlusion of cerebral vessels.

In case that temporary cerebral ischemia occurs, the supply of oxygen and glucose to brain is prevented and several syndrome such as ATP decrease and edema follows, which causes to exclusive range of brain injury as the result. After the considerable time lapses, the apoptosis of neuronal cell occurs, which is called as delayed neuronal death. The effect on the delayed type of neuronal death is performed by transient forebrain ischemic model using Mongolian gerbil and it has been reported that neuronal death occurs at CA1 region in hippocampus four days after the inducement of cerebral ischemia for 5 mins (Kirino T et al, Acta Neuropathol., 62 pp 201-208, 1984; Kirino T, Brain Res., 239, pp 57-69, 1982).

It has been reported that the neuronal death is caused by two mechanism; one is excitotoxic neuronal death mechanism; cerebral ischemia causes to excessive accumulation of outer glutamate and those glutamate are influxed into inner cell which causes to neuronal death by excessive accumulation of intracellular calcium ion (Kang T. C. et al, J. Neurocytol., 30, pp 945-955, 2001) and another is oxidative neuronal death; the ischemia--repurfusion causing abrupt supply of oxygen results in the increase of free radical to give rise to the injury of DNA and cytoplasm (Won M. H. et al, Brain Res., 836, pp 70-78, 1999; Sun A. Y. et al, J. Biomed. Sci., 5, pp 401-141, 1998; Flowers F et al, New Horiz, 6, pp 169-180, 1998).

On the base of those mechanism study, there have been endeavored to develop effectively inhibiting substance of neuronal death or the mechanism thereof till now, however, neuroprotective agents have been nearly not yet found.

t-PA (tissue plasminogen activator), sole approved cerebral ischemia treating agent in FDA and sold in the market is a thrombolytic agent dissolving thrombus causing cerebral ischemia and inducing rapid supply of oxygen and glucose. Accordingly, it could not protect neuronal cell directly therefore it should be used urgently. Furthermore, since it is a thrombolytic agent, hemorrhagic cerebral disease occurs in case that it is administrated in over dose or too frequently.

MK-801, a potent NMDA receptor blocker effectively inhibiting initial calcium ion influx had been on clinical trial however it was abandoned because of its adverse action.

In Korea, lots of health care foods containing natural substance have been on the market however most of those are not yet authorized by scientific test and abused to give rise to scientific problems in the end.

Accordingly, it have been still needed to develop novel natural resource effective in treating and preventing cerebral disease through substantive and scientific experiments till now.

Panax ginseng C. A. Meyer belonged to Araliaceae, is distributed in Asian countries as well as other countries. It has been used as materials of Chinese medicine as a restorative and reported to comprise various ginseng saponins such as ginsenoside Rb1, Rb2, Rc, Rd, Re, panaxadiol, panaxatriol, kaempferol etc (Chung B. S. and Shin M. K.; HyangyakDaesacheon, Youngrimsa., pp 439-444, 1998).

Acanthopanax senticosus HARMS belonged to Araliaceae, is distributed in humid area of Korea, Siberia etc. It has been used as materials of Chinese medicine as adatogenic activities, anti-inflammatory agents and reported to comprise various glycosides such as saponins, eleutherosides, essential oil etc (Chung B. S. and Shin M. K.; HyangyakDaesacheon, Youngrimsa., pp 433-435, 1998).

Angelica sinensis DIELS belonged to Umbelliferae, is distributed in China. It has been used as materials of Chinese medicine as a treating agent of various gynopathic diseases and reported to comprise various components such as ligustilide, butylidene phthalide, cnilide, isocnilide, nicotinic acid, sitosterol, essential oil etc (Chung B. S. and Shin M. K.; HyangyakDaesacheon, Youngrimsa., pp 406-408, 1998).

Scutellaria baicalensis GEORGI belonged to Labiatae, is distributed in Korea, China, Siberia etc. It has been used as materials of Chinese medicine as a diuretics, anti-inflammatory agent, cholagogue, etc and reported to comprise various components such as baicalin, baicalin, woginin, wogonoside, scutellarein, hispidulin etc (Chung B. S. and Shin M. K.; HyangyakDaesacheon, Youngrimsa., pp 864-865, 1998).

However, there has been not reported or disclosed about therapeutic effect for brain disease of the crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI, in any of above cited literatures, the disclosures of which are incorporated herein by reference.

To investigate an effect of the extract of above described crude drug complex on the neuronal growth and differentiation through several biochemical experiments and to confirm whether the extract play an important role in inhibiting neuronal apoptosis, main cause of stroke and neurodegenerative diseases, the inventors of the present invention have intensively carried out 4 sensory motor function tests using animal model, and finally completed present invention by confirming that the extract inhibit the neuronal apoptosis, and show neuroprotective activity.

These and other objects of the present invention will become apparent from the detailed disclosure of the present invention provided hereinafter.

DISCLOSURE OF INVENTION

Technical Problem

In case that temporary cerebral ischemia occurs, the supply of oxygen and glucose to brain is prevented and several syndrome such as ATP decrease and edema follows, which causes to exclusive range of brain injury as the result. After the considerable time lapses, the apoptosis of neuronal cell occurs, which is called as delayed neuronal death. The effect on the delayed type of neuronal death is performed by transient forebrain ischemic model using Mongolian gerbil and it has been reported that neuronal death occurs at CA1 region in hippocampus four days after the inducement of cerebral ischemia for 5 mins (Kirino T et al, Acta Neuropathol., 62 pp 201-208, 1984; Kirino T, Brain Res., 239, pp 57-69, 1982).

It has been reported that the neuronal death is caused by two mechanism; one is excitotoxic neuronal death mechanism; cerebral ischemia causes to excessive accumulation of outer glutamate and those glutamate are influxed into inner cell which causes to neuronal death by excessive accumulation of intracellular calcium ion (Kang T. C. et al, J. Neurocytol., 30, pp 945-955, 2001) and another is oxidative neuronal death; the ischemia--repurfusion causing abrupt supply of oxygen results in the increase of free radical to give rise to the injury of DNA and cytoplasm (Won M. H. et al, Brain Res., 836, pp 70-78, 1999; Sun A. Y. et al, J. Biomed. Sci., 5, pp 401-141, 1998; Flowers F et al, New Horiz, 6, pp 169-180, 1998).

On the base of those mechanism study, there have been endeavored to develop effectively inhibiting substance of neuronal death or the mechanism thereof till now, however, neuroprotective agents have been nearly not yet found.

t-PA (tissue plasminogen activator), sole approved cerebral ischemia treating agent in FDA and sold in the market is a thrombolytic agent dissolving thrombus causing cerebral ischemia and inducing rapid supply of oxygen and glucose. Accordingly, it could not protect neuronal cell directly therefore it should be used urgently. Furthermore, since it is a thrombolytic agent, hemorrhagic cerebral disease occurs in case that it is administrated in over dose or too frequently.

MK-801, a potent NMDA receptor blocker effectively inhibiting initial calcium ion influx had been on clinical trial however it was abandoned because of its adverse action.

In Korea, lots of health care foods containing natural substance have been on the market however most of those are not yet authorized by scientific test and abused to give rise to scientific problems in the end.

Technical Solution

Accordingly, it is an object of the present invention to provide a pharmaceutical composition comprising an extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI as an active ingredients for the treatment and prevention of stroke and neurodegenerative diseases by protecting neuronal cell.

Preferably, above described extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI is mixed in the ratio of 1-5:2-7:2-7:1, more preferably, 2:5:4:1.

Above described extract comprises the extract prepared by extracting plant material with water, lower alcohols such as methanol, ethanol, preferably methanol, or the mixtures thereof.

It is an object of the present invention to provide a use of an extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI for the preparation of therapeutic agent for the treatment and prevention of stroke and neurodegenerative diseases by protecting neuronal cell in human or mammal in need thereof.

It is an object of the present invention to provide a method of treating or preventing stroke and neurodegenerative diseases by protecting neuronal cell in a mammal comprising administering to said mammal an effective amount of an extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI, together with a pharmaceutically acceptable carrier thereof.

It is another object of the present invention to provide a health food or food additives comprising an extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI, together with a sitologically acceptable additive for the prevention and improvement of stroke and neurodegenerative diseases by protecting neuronal cell.

Above described neurodegenerative diseases comprises apoplexy, Alzheimer's disease (AD), Parkinson's disease (PD), Pick's disease, Creutzfeldt-Jakob disease, senile dementia and the like.

The pharmaceutical composition of the present invention can contain about 0.01.about.50% by weight of the above extract based on the total weight of the composition.

The health food of the present invention comprises above extracts as 0.01 to 80%, preferably 1 to 50% by weight based on the total weight of the composition.

Above health food can be contained in health food, health beverage etc, and may be used as powder, granule, tablet, chewing tablet, capsule, beverage etc.

An inventive extract may be prepared in accordance with the following preferred embodiment.

Hereinafter, the present invention is described in detail.

An inventive extract of the present invention can be prepared in detail by following procedures,

The inventive crude extract of crude drug complex comprising Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI, can be prepared by follows; all the dried materials excepting Scutellaria baicalensis GEORGI is dried, cut, crushed and mixed with 5 to 25-fold, preferably, approximately 10 fold volume of distilled water, lower alcohols such as methanol, ethanol, butanol and the like, or the mixtures thereof, preferably hot water; the solution is treated with hot water at the temperature ranging from 20 to 100.degree. C., preferably from 60 to 100.degree. C., for the period ranging from 1 to 24 hrs with extraction method by the extraction with hot water, cold water, reflux extraction, or ultrasonication extraction with 1 to 5 times, preferably 2 to 3 times, consecutively; the residue is added to honey and remaining dried Scutellaria baicalensis GEORGI to be subjected to further extraction with hot water for the period ranging from 48 to 120 hrs, preferably, 96 hrs; and the colleted extract is filtered to obtain the supernatant to be concentrated with rotary evaporator at the temperature ranging from 20 to 100.degree. C., preferably from 50 to 70.degree. C. and then dried by vacuum freeze-drying, hot air-drying or spray drying to obtain dried extract powder of the crude drug complex which can be soluble in water, lower alcohol, or the mixtures thereof.

In an alternative embodiment of the present invention, the extract of crude drug of the present invention can be prepared by the procedure comprising the steps consisting of; (a), adding dried Panax ginseng C. A. Meyer and Acanthopanax senticosus HARMS to 5 to 25-fold, preferably, 2 to 8-fold volume of distilled water, lower alcohols such as methanol, ethanol, butanol and the like, or the mixtures thereof, preferably hot water and extracting at the temperature ranging from 20 to 100.degree. C., preferably from 60 to 100.degree. C., for the period ranging from 1 to 24 hrs with reflux extraction to collect their extract, filtering and drying to obtain concentrated extract; (b) adding the mixture consisting of the extract of Poria cocos WOLF and Rehmania gluticosa L, and honey thereto and extracting with hot water for the period ranging from 48 to 120 hrs, preferably 96 hrs with the addition of the extract of Scutellaria baicalensis GEORGI to obtain crude complex drug extract of the present invention.

Accordingly, present invention also provides the extract of crude drug complex prepared by above described preparation method.

Additionally, polar solvent soluble and non-polar solvent soluble extract of present invention can be prepared by following procedure; the crude extract prepared by above step is suspended in water, and then is mixed with 1 to 100-fold, preferably, 1 to 5-fold volume of non polar solvent such as ethyl acetate, chloroform, hexane and the like; the non-polar solvent soluble layer is collected to obtain non-polar solvent soluble extract of the present invention and remaining polar solvent soluble layer is collected to obtain polar solvent soluble extract of the present invention which is soluble in water, lower alcohol, or the mixture thereof. Also, above described procedures may be modified or subjected to further step to fractionate or isolate more potent fractions or compounds by conventional procedure well-known in the art, for example, the procedure disclosed in the literature (Harborne J. B. Phytochemical methods: A guide to modern techniques of plant analysis, 3.sup.rd Ed. pp 6-7, 1998).

Through the experiment to investigate the effect of the extract of above described crude drug complex on neuronal injury caused by regional cerebral ischemia occurred in MCAo animal model, it is confirmed that the extract inhibits the neuronal apoptosis caused by regional cerebral ischemia and shows neuroprotective activity.

In accordance with another aspect of the present invention, there is provided a pharmaceutical composition comprising the extract of above described crude drug complex prepared by above described preparation method for the treatment and prevention of stroke and neurodegenerative diseases by protecting neuronal cell as active ingredients.

It is another of the present invention to provide method of treating or preventing stroke and neurodegenerative diseases by protecting neuronal cell in a mammal comprising administering to said mammal an effective amount of an extract of above described crude drug complex prepared by above preparation method.

The inventive composition for treating and preventing stroke and neurodegenerative diseases by protecting neuronal cell, may comprises above extracts as 0.01.about.50% by weight based on the total weight of the composition.

The inventive composition may additionally comprise conventional carrier, adjuvants or diluents in accordance with a using method well known in the art. It is preferable that said carrier is used as appropriate substance according to the usage and application method, but it is not limited. Appropriate diluents are listed in the written text of Remington's Pharmaceutical Science (Mack Publishing co, Easton Pa.).

Hereinafter, the following formulation methods and excipients are merely exemplary and in no way limit the invention.

The composition according to the present invention can be provided as a pharmaceutical composition containing pharmaceutically acceptable carriers, adjuvants or diluents, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starches, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil. The formulations may additionally include fillers, anti-agglutinating agents, lubricating agents, wetting agents, flavoring agents, emulsifiers, preservatives and the like. The compositions of the invention may be formulated so as to provide quick, sustained or delayed release of the active ingredient after their administration to a patient by employing any of the procedures well known in the art.

For example, the compositions of the present invention can be dissolved in oils, propylene glycol or other solvents that are commonly used to produce an injection. Suitable examples of the carriers include physiological saline, polyethylene glycol, ethanol, vegetable oils, isopropyl myristate, etc., but are not limited to them. For topical administration, the extract of the present invention can be formulated in the form of ointments and creams.

Pharmaceutical formulations containing present composition may be prepared in any form, such as oral dosage form (powder, tablet, capsule, soft capsule, aqueous medicine, syrup, elixirs pill, powder, sachet, granule), or topical preparation (cream, ointment, lotion, gel, balm, patch, paste, spray solution, aerosol and the like), or injectable preparation (solution, suspension, emulsion).

The composition of the present invention in pharmaceutical dosage forms may be used in the form of their pharmaceutically acceptable salts, and also may be used alone or in appropriate association, as well as in combination with other pharmaceutically active compounds.

The desirable dose of the inventive extract or composition varies depending on the condition and the weight of the subject, severity, drug form, route and period of administration, and may be chosen by those skilled in the art. However, in order to obtain desirable effects, it is generally recommended to administer at the amount ranging 10 g/kg, preferably, 1 to 3 g/kg by weight/day of the inventive extract of the present invention. The dose may be administered in single or divided into several times per day. In terms of composition, the amount of inventive extract should be present between 0.01 to 50% by weight, preferably 0.5 to 40% by weight based on the total weight of the composition.

The pharmaceutical composition of present invention can be administered to a subject animal such as mammals (rat, mouse, domestic animals or human) via various routes. All routes of administration are contemplated, for example, administration can be made orally, rectally or by intravenous, intramuscular, subcutaneous, intracutaneous, intrathecal, epidural or intracerebroventricular injection.

Also, the present invention provide a health food comprising an extract of crude drug complex consisting of Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS, Scutellaria baicalensis GEORGI, together with a sitologically acceptable additive for the prevention and improvement of stroke and neurodegenerative diseases by protecting neuronal cell. The health food can be prepared by adding 0.01 to 80 w % of above described extracts, amino acids 0.001 to 5% by weight, vitamins 0.001 to 2% by weight, sugars 0.001 to 20% by weight, organic acids 0.001 to 10% by weight and proper amount of sweetener and flavors sitologically acceptable food additive.

Above described extract of above described crude drug complex can be added to food and beverage for the prevention and improvement of stroke and neurodegenerative diseases by protecting neuronal cell.

To develop for health food, examples of addable food comprising above extracts of the present invention are various food, beverage, gum, vitamin complex, health improving food and the like, and can be used as power, granule, tablet, chewing tablet, capsule or beverage etc.

Also, the extract of the present invention could prevent and improve stroke and neurodegenerative diseases by protecting neuronal cell by comprising to child and infant food, such as modified milk powder, modified milk powder for growth period, modified food for growth period.

Above described composition therein can be added to food, additive or beverage, wherein, the amount of above described extract in food or beverage may generally range from about 0.1 to 80 w/w %, preferably 1 to 50 w/w % of total weight of food for the health food composition and 1 to 30 g, preferably 3 to 10 g on the ratio of 100 of the health beverage composition.

Providing that the health beverage composition of present invention contains above described extract as an essential component in the indicated ratio, there is no particular limitation on the other liquid component, wherein the other component can be various deodorant or natural carbohydrate etc such as conventional beverage. Examples of aforementioned natural carbohydrate are monosaccharide such as glucose, fructose etc; disaccharide such as maltose, sucrose etc; conventional sugar such as dextrin, cyclodextrin; and sugar alcohol such as xylitol, and erythritol etc. As the other deodorant than aforementioned ones, natural deodorant such as taumatin, stevia extract such as levaudioside A, glycyrrhizin et al., and synthetic deodorant such as saccharin, aspartam et al., may be useful favorably. The amount of above described natural carbohydrate is generally ranges from about 1 to 20 g, preferably 5 to 12 g in the ratio of 100 of present beverage composition.

The other components than aforementioned composition are various nutrients, a vitamin, a mineral or an electrolyte, synthetic flavoring agent, a coloring agent and improving agent in case of cheese chocolate et al., pectic acid and the salt thereof, alginic acid and the salt thereof, organic acid, protective colloidal adhesive, pH controlling agent, stabilizer, a preservative, glycerin, alcohol, carbonizing agent used in carbonate beverage et al. The other component than aforementioned ones may be fruit juice for preparing natural fruit juice, fruit juice beverage and vegetable beverage, wherein the component can be used independently or in combination. The ratio of the components is not so important but is generally range from about 0 to 20 w/w % per 100 w/w % present composition. Examples of addable food comprising aforementioned extract therein are various food, beverage, gum, vitamin complex, health improving food and the like.

The inventive composition may additionally comprise one or more than one of organic acid, such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid; phosphate, such as phosphate, sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate; natural anti-oxidants, such as polyphenol, catechin, .alpha.-tocopherol, rosemary extract, vitamin C, green tea extract, licorice root extract, chitosan, tannic acid, phytic acid etc.

The above-described extract of the present invention may be 20 to 90% high concentrated liquid, power, or granule type.

Similarly, the above-described extract of the present invention can additionally comprise at least one lactose, casein, dextrose, glucose, sucrose and sorbitol.

Inventive extract of the present invention have no toxicity and adverse effect therefore; they can be used with safe.

It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions, use and preparations of the present invention without departing from the spirit or scope of the invention.

Claim 1 of 5 Claims

1. A pharmaceutical composition comprising an effective amount of an extract of crude drug complex wherein, the crude drug complex comprises Panax ginseng C. A. Meyer, Acanthopanax senticosus HARMS, Angelica sinensis DIELS and Scutellaria baicalensis GEORGI as active ingredients, for treating and/or reducing the risk of a stroke caused by cerebral ischemia.
 

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