WO2005011719A1

WO2005011719A1 - Composition for preventing or treating dementia comprising acanthopanax extract

Info

Publication number: WO2005011719A1
Application number: PCT/KR2004/001929
Authority: WO
Inventors: Kwang Soo Soung; Dong Keun Song
Original assignee: Biosynergen, Inc.
Priority date: 2003-07-30
Filing date: 2004-07-30
Publication date: 2005-02-10
Also published as: KR20050014710A

Abstract

The present invention relates to a composition for preventing or treating dementia comprising acanthopanax extract. Acanthopanax extract according to the present invention is extracted by adding water, distilled water, alcohol, nucleic acid, ethyl acetate, acetone, chloroform, methylene chloride, or mixtures thereof.

Description

COMPOSITION FOR PREVENTING OR TREATING DEMENTIA COMPRISING ACANTHOPANAX EXTRACT

Technical Field The present invention relates to a composition for preventing or treating dementia comprising acanthopanax extract.

Background Art Along with the increase in the aged population worldwide as well as in Korea, all kinds of degenerative geriatric disease that are common in aged people have caused social and financial losses. Especially, a degenerative brain disorder such as Alzheimer's disease is known that persons under the age of 40 are hardly affected thereby, however, persons over the age of 60 are affected thereby twofold every five year, and in persons over the age of 85, that number rises to 40-50%. Alzheimer Association of U.S.A. estimates that the yearly financial loss in dementia disease is approximately 10 million dollars. Presumably, among the aged population under the age of 65, about 245,000 people are affected by dementia every year and approximately 14 million people suffer from dementia. And furthermore, its death rate is very high, just next to cardiovascular disease, a malignant tumor, and apoplexy. Although it is difficult to estimate the numerical value precisely due to lack of the domestic reference data, more than 300,000 people are presumed to be dementia patients under the estimate that approximately 4-5% of the aged population over the age of 65 is suffering from dementia. Since dementia is incurable disease which not only devastates a patient's human living, but also destroys family members' living, the disease is making its appearance as a serious social and financial problem. Accordingly, although there are many studies being made for preventing and treating dementia, there are still plenty of difficulties lying in developing treatment medicine since the cause of dementia disease is not revealed. Most treatments of dementia are presently focused on restraining impairment of one's memory that appears in the first stage of dementia. Also, since new substance therapeutics has high possibility that can be found from natural substances that have long been adapted through a folk remedy, there are active attempts in progress to develop new medicine from natural substances, especially, herbal medicine. On the one hand, acanthopanax is used as medicine in herbal treatment. Acanthopanax tastes pungent and is worm-natured. It is known that acanthopanax acts on liver-ligaments and nerves to remove joint-ache, to cheer one up and to excite the spirits. It is also known that acanthopanax replenishes 'Oro' (a physical and mental fatigue which results from the weakness of the five viscera) and ^Chilsang' (seven symptoms which appear when a man is in poor health) and is used for being worked off one's legs. Further, if acanthopanax is taken for a long time, it is known that acanthopanax heightens '-7/ ' of the body, protects the stomach, invigorates, refreshes the spirits, improves willpower, lightens the body, prevents aging, makes the bad blood inside the body clean and maintains the same clean. Furthermore, it is known that acanthopanax cures various kinds of symptoms such as pain in the waist and the backbone, impotence, testis sweating, vulvar itching, etc. Accordingly, there are presently studies about various physiological activities of acanthopanax in progress. The present invention has been accomplished after finding that acanthopanax is effective in improving memory loss as well as not toxic during making a progress of the study about various physiological activities of acanthopanax.

Disclosure of the Invention Technical Question The object of the present invention is to provide a composition for preventing or treating dementia comprising acanthopanax extract.

Technical Solution The present invention provides a composition for preventing or treating dementia comprising acanthopanax extract as an effective ingredient. As confirmed in <Preparations> and <Test Example> below, when the present inventors extracted acanthopanax divaricatus or acanthopanax senticosus through distilled water, ethanol aqueous solution, or methanol aqueous solution, and then executed a passive avoidance test with the extract, the inventors could confirm that passive avoidance reaction time has decreased in a group wherein only beta-amyloid that was known as a causative factor of dementia such as Alzheimer's disease (Selkoe, Annu. Rev. Neurosci., 17, 489-517(1994); it is regarded that the content of this record is included in the present specification) was prepared, however, that passive avoidance reaction time has increased in a group wherein beta-amyloid was prepared along with acanthopanax extract that was extracted through <Test Example> of the present invention. The passive avoidance test is a test using mice for measuring memory ability thereof about unpleasant stimuli for 24 hours. In this passive avoidance test, decrease of passive avoidance reaction time means that memory ability of a test-subjected mouse about the unpleasant stimuli has decreased. On the other hand, increase of passive avoidance reaction time means that memory ability of a test-subjected mouse about the unpleasant stimuli has increased. Furthermore, it was shown that acanthopanax extract from <Preparations> below of the present invention has decreased the density of malonaldehyde which is known as an index of oxidative stress acting on the outbreak of a degenerative neurogical disorder, and has increased the density of acetylcholine, a transmitting substance of brain nerve, whose density is known to be decreased among dementia patients, in the test using mice. Like this, the result that passive avoidance reaction time has increased or the result that the density of malonaldehyde has decreased and the density of acetylcholine has increased through the acanthopanax extract may show that the acanthopanax extract is effective in preventing or treating dementia such as Alzheimer's disease, etc. In one embodiment, the present invention provides a pharmaceutical composition for preventing or treating dementia comprising acanthopanax extract as an effective ingredient. In implementing a clinical administration, the pharmaceutical composition of the invention can be administered via various routes including an oral route, a percutaneous route, a subcutaneous route, an intravenous route, and an intramuscular route. Furthermore, it is possible to formulate a pharmaceutical composition of the invention into various shapes of medicine. In this case, it can be formulated by using flocculating agents or diluents such as a filler, an increasing agent, a bonding agent, a wetting agent, and a surface active agent. Solid types for an oral administration include tablets, pills, powders, granule, and capsules, etc, wherein the solid types are compounded with mixtures of at least one flocculating agents such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to these flocculating agents, lubricants also can be used. Liquid types for an oral administration comprise suspension, liquids, emulsions, syrup, etc, wherein liquid types can include various flocculating agents such as a wetting agent, a flavor, an aromatic, a storing agent, etc besides diluents such as water and liquid paraffin. Types for non-oral administration include a sterilized aqueous solution, non-aqueous solvent, suspension, emulsion, lyophilization, and a suppository. It can be used propylene glycol, polyethylene glycol, vegetable oil such as olive oil and ester injectable like ethyl oleate for non-aqueous solvent and suspension. It can be used witepsol, macrogol, tween 61 , cacao butter, laurin butter, glycerol and gelatin for a suppository. The pharmaceutical composition of the invention can be administered at a dose of about 1 to 30 mg/kg of body weight per day and the pharmaceutical composition can be administered totally once or divisionally several times a day. It should be understood, however, that the specific dose level of the pharmaceutical composition of the invention will depend on the variety of factors such as a route of administration, the patient age, sex, body weight, severity of the disease, etc, and thus, the above dose level does not limit the scope of the present invention in any aspect. In other embodiments, the present invention provides a food composition for preventing or treating dementia comprising acanthopanax extract as an effective ingredient. The food composition of the present invention can be formulated into the shapes of gums, vitamin compounds, health aid food, special nutrition food, functional drinkables, etc. Besides the fact that the food composition comprises acanthopanax extract as an effective ingredient, it is possible to add monosaccharide such as glucose and fructose, disaccharide such as maltose and sucrose, or, polysaccharide such as dextrin and cyclo-dextrin thereto. Furthermore, it is possible to add other food additive including sugar alcohol such as xylitol, sorbitol, erythritol, etc and a flavor such as taurine, stevia extract, etc. In the meantime, as shown in <Test Example> below, there was no evidence of significant toxicity at the result of an acute toxicity test using mice of acanthopanax extract. In the present invention, it is that the acanthopanax extract comprised in the composition for preventing or treating dementia as an effective ingredient is extracted by adding water, distilled water, alcohol, nucleic acid, ethyl acetate, acetone, chloroform, methylene chloride or mixtures thereof into acanthopanax. And it is more preferable that the acanthopanax extract is extracted by adding water, distilled water, alcohol, or, alcohol aqueous solution into the acanthopanax. Herein, it is preferable that the above alcohol is low-grade alcohol in the aspect of efficiency of extract, wherein low-grade alcohol includes methanol, ethanol, or butanol. Furthermore, although it was used acanthopanax senticosus and acanthopanax divaricatus in <Preparations> below, it may be used other kinds of acanthopanax. More concretely, the acanthopanax extract which is comprised in the composition for preventing or treating dementia as an effective ingredient is one of those described below: (a) acanthopanax extract which is prepared by extracting acanthopanax divaricatus or acanthopanax senticosus using distilled water; (b) acanthopanax extract which is prepared by extracting acanthopanax divaricatus or acanthopanax senticosus using ethanol aqueous solution; and (c) acanthopanax extract which is prepared by extracting acanthopanax divaricatus or acanthopanax senticosus using methanol aqueous solution. More concretely, the acanthopanax extract is one of those described below: (i) acanthopanax extract which is prepared by boiling-extracting acanthopanax divaricatus or acanthopanax senticosus using distilled water; (ii) acanthopanax extract which is prepared by stirring acanthopanax divaricatus or acanthopanax senticosus with ethanol aqueous solution; and (iii) acanthopanax extract which is prepared by stirring acanthopanax divaricatus or acanthopanax senticosus with methanol aqueous solution. How long to extract by boiling water or how long to extract by stirring should be within the scope of the contents (Preparation 1 to 6 below) of the present specification and within the scope of general ability of those skilled in the art, as far as it is based on the preceding art of the time when the present invention was applied. Brief description of the Drawings Fig. 1 is a graph showing the result of passive avoidance reaction of mice administered with acanthopanax extract. Fig. 2 is a graph showing the decrease of density of malonaldehyde of mice administered with acanthopanax extract. Fig. 3 is a graph showing the increase of density of acetylcholine of mice administered with acanthopanax extract.

Best Mode for Carrying out the Invention Hereinafter, the present invention will be in detail described with reference to the embodiments. However, it is very evident for those skilled in the art that these embodiments or the like do not limit the scope of the present invention. <Preparations> Preparation of Acanthopanax Extract

Preparation Example 1> Preparation! of Acanthopanax Extract 200g of acanthopanax divaricatus was placed in 3 liter plastic and boiling- extracted for 16 hours after adding 1500ml distilled water. The aimed acanthopanax extract was prepared by decompressing and concentrating the obtained extract in vacuous state, after filtering the obtained solution by using a filter paper.

Preparation Example 2> Preparation 2 of Acanthopanax Extract Acanthopanax extract was prepared under the same conditions and processes as the above Preparation Example 1> using acanthopanax senticosus instead of acanthopanax divaricatus.

Preparation Example 3> Preparation 3 of Acanthopanax Extract 200g of acanthopanax divaricatus was placed in 3 liter plastic and stirred for 2 days at the state of room temperature after adding 1500ml ethanol aqueous solution of 70%. The aimed acanthopanax extract was prepared by lyophilizing the obtained extract, after filtering the obtained solution by using a filter paper. Preparation Example 4> Preparation 4 of Acanthopanax Extract Acanthopanax extract was prepared under the same conditions and processes as the above Preparation Example 3> using acanthopanax senticosus instead of acanthopanax divaricatus.

Preparation Example 5> Preparation 5 of Acanthopanax Extract Acanthopanax extract was prepared under the same conditions and processes as the above Preparation Example 3> using methanol aqueous solution of 85 % instead of ethanol aqueous solution as extract solution.

Preparation Example 6> Preparation 6 of Acanthopanax Extract Acanthopanax extract was prepared under the same conditions and processes as the above Preparation Example 3> using methanol aqueous solution of 85 % instead of ethanol aqueous solution as extract solution and using acanthopanax senticosus instead of acanthopanax divaricatus

<Embodiments> Confirmation of the effect for preventing or treating dementia and toxicity test of acanthopanax extract

<Embodiment Example 1> Embodiment of a Passive Avoidance Test A passive avoidance test was implemented in order to confirm effects for improvement of memory ability of acanthopanax extract prepared from Preparation 1>. In this test, 30 mice at the age of 4-5 week and weight of 24~28g were divided into 3 groups of each 10 mice. For mice of one test group among three groups, acanthopanax extract prepared from Preparation Example 1> was dissolved in water at the density of 0.5g/kg and then orally administered via oral syringe for mouse five times respectively at intervals of 12 hours for 3 days. Consequently, acanthopanax extract of Preparation 1> was administered in the amount of 0.25g/kg. And normal sodium chloride was administered to mice of other two test groups five times respectively for 3days. After that, each 1.85 μg of beta-amyloid (1 -42) (A Bι. ₂) was intracerebroventricularly injected into each mouse of one test group administered with acanthopanax extract and of one of two groups administered only by sodium chloride, 19 hours after starting administration of acanthopanax extract and sodium chloride. Intracerebroventricular injection followed the method described in Laursen and Belknap (Laursen and Belknap, J. Pharmacol, Methods 16:355-357 (1986)). That is, beta-amyloid was injected intracerebroventricularly by using a 50 μi Hamilton Syringe fitted with a 26-guage needle until the tip of the needle was inserted 2.4mm deep into the bregma of each mouse. On days 1-2 after the intracerebroventricular injection, the passive avoidance test was implemented according to the method described in Song (Song et al, J. Neurochem., 71 , 875-878(1998)). In the above test, it was used a passive avoidance chamber consisting of an illuminated compartment and a dark compartment, in which means to deliver electrical shock is constructed on the bottom thereof. First, a mouse was placed into the illuminated compartment, and then as soon as the mouse entered the dark room, an electric shock (0.25 mA) was immediately applied to the mouse for one second. After 24 hours of training, the mouse was again placed in the illuminated compartment and the latency to enter the dark compartment, that is, passive avoidance reaction time, was measured. The maximum limit of time was set to 300 seconds. If the mouse did not enter the dark compartment even after 300 seconds, the latency of 300 seconds was determined as the passive avoidance reaction time. The results of the passive avoidance test above are shown in Fig. 1, expressed with the average value of latencies of 10 mice. As confirmed in Fig. 1, while the mice in the test group administered with beta- amyloid (1-42) (A β ι-₄₂) showed significant decrease in passive avoidance reaction time compared with those in the sham group (administered only by sodium chloride), the mice in the test group administered with acanthopanax extract of Preparation Example 1> showed significant increase in passive avoidance reaction time compared with the group administered with beta-amyloid (1 -42) (A Bi-₄₂). From this, it can be understood that acanthopanax extract can improve declined memory and, consequently, can prevent and treat dementia. In the mean time, in case of acanthopanax extract of <Preparation Example 2> to Preparation Example 6>, when the tests were implemented under the same conditions and progresses as the above <Embodiment 1>, although the mice in the tests showed a little deviation compared with the mice administered with acanthopanax extract of Preparation Example 1>, the mice also showed increase in passive avoidance reaction time, just like acanthopanax extract of Preparation Example 1> (not showing the results).

<Embodiment Example 2> Measurement of density of malonaldehyde Peroxide acts on oxidation substance such as malonaldehyde (hereinafter, "MAD") or the like to increase and be stored in the body. It is known that oxidation substance such as MAD or the like causes inflammatory disease such as arthritis, arteriosclerosis, dementia, etc. It was observed how the amount of MAD known to be related with disease such as dementia would be varied by acanthopanax extract prepared from Preparation Example 1>. In this test, 30 mice at the age of 4-5 week and weight of 24~28g were divided into 3 groups of each 10 mice. For mice of one test group among three groups, acanthopanax extract prepared from Preparation Example 1> was orally administered via oral syringe for mouse at the density of 0.25g/kg and for mice of other two test groups, sodium chloride was administered in the amount of 0.25g/kg. After that, each

1.85 βg of beta-amyloid (1 -42) (A βι-4₂) was intracerebroventricularly injected into each mouse of one test group administered with acanthopanax extract and of one of two groups administered only by sodium chloride. On days 5 after intracerebroventricular injection, the predetermined region of cerebral cortex was stored in the frozen state of -

80 ^°C after the picking thereof. MAD density of the inside brain was measured by using

Bioxytech LPO-586 kit (OXIS international. Inc, U.S.A). This is to pick a portion of the brain tissue and then add 20 mM tris-HCL (pH 7.4) solution thereto and homogenize the same and finally pick the upper portion through centrifugal separator. Density of MAD was measured by measuring absorbance at the wavelength of 586mm of MAD- derivative which had been produced after adding reaction solution (N-methyl-2- phenylindole 650 μi, ethanesulfonic acid 150 μJL) to the upper portion and then culturing the added upper portion at the temperature of 45 ^°C for 60 minutes. The results are shown in Fig. 2, expressed with the average value of latencies of 10 mice. As confirmed in Fig. 2, it was observed that the mice in the test group administered with beta-amyloid (1-42) (A βι-₄₂) showed significant increase in MAD density compared with those in the sham group and the mice in the test group administered with acanthopanax extract showed decrease in MAD density. In the mean time, in case of acanthopanax extract of Preparation Example 2> to Preparation Example 6>, although the mice in the tests showed a little deviation compared with the mice administered with acanthopanax extract of Preparation Example 1>, the mice also showed decrease in MAD density, just like acanthopanax extract of Preparation Example 1> (not showing the results).

<Embodiment Example 3> Measurement of density of acetylcholine In case of dementia patients of Alzheimer's disease type, it is known that drastic decrease of density of acetylcholine causes memory and cognitive ability disorder, wherein acetylcholine is a nerve transmitting substance. In this test, 30 mice at the age of 4-5 week and weight of 24~28g were divided into 3 groups of each 10 mice. For mice of one test group among three groups, acanthopanax extract prepared from Preparation Example 1> was orally administered via oral syringe for mouse at the density of 0.25g/kg and for mice of other two test groups, sodium chloride was administered in the amount of 0.25g/kg. After that, each 1.85 μg of beta-amyloid (1-42) (A βι-₄₂) was intracerebroventricularly injected into each mouse of one test group administered with acanthopanax extract and of one of two groups administered only by sodium chloride. On days 5 after intracerebroventricular injection, the predetermined region of cerebral cortex was stored in the frozen state of - 80 ^°C after the picking thereof. The amount of acetylcholine followed the method described in chemi- luminescent method of J.Neurohem, 1981, 37, 1475 and Neurochem Int, 1981, 3, 81. After extracting protein from the brain by using 5%-trichloroacatic acid, it is to eliminate trichloroacatic acid by washing the extracted protein with ether. It is to dissolve the remaining protein produced by evaporating remaining of ether into sodium hydroxide of IM and then put buffer solution therein and neutralize it. And then, it is to mix exhibition of 50 C, with reaction buffer equally (mixtures of 250units/ml choline oxidase 100 μJl., 2mg/ml-peroxidae 50 μi and 1 mM-Luminol 100/-!). Continuously, it is to hydrolyze acetylcholine with acetylcholine s-lase of 5 μi.. And then it was measured volume of luminous light by using luminometer (Luminometer: Lumat LB 9507). The results of the above test are shown in Fig. 3, expressed with the average value of latencies of 10 mice. As confirmed in Fig. 3, the mice in the test group administered with beta- amyloid showed decrease in acetylcholine density compared with those in the sham group, which is identical to the already known result that dementia patients of Alzheimer's disease type have shown decrease in acetylcholine density, a nerve transmitting substance. And it was observed that the mice in the test group administered with acanthopanax extract showed decrease in acetylcholine density compared with those in the test group administered with beta-amyloid. In the mean time, in case of acanthopanax extract of Preparation Example 2> to Preparation Example 6>, although the mice in the tests showed a little deviation compared with the mice administered with acanthopanax extract of Preparation

Example 1>, the mice also showed decrease in acetylcholine density, just like acanthopanax extract of <Preparation Example 1> (not showing the results).

<Test Example> Acute toxicity test 6 mice at the age of 4-5 week and weight of 24~28g were divided into 2 groups of each 3 mice. For mice of one test group, acanthopanax extract of Preparation Example 1> was orally administered via oral syringe for mouse once in the amount of lg/kg/ respectively. For 7days after administration, it was observed a change in general symptoms, outbreak of dead animals and a change in body weight. In addition, it was implemented hematological test and hemo-biochemistric test. The result revealed no typical clinical symptoms in any groups of mice administered with acanthopanax extract and any toxicity was observed in aspect of a change in body weight, hematological test and hemo-biochemistric test for mice of the groups. Furthermore, acanthopanax extract showed no toxicity in any group of mice up to 60g/kg and was decided as a safe substance, wherein the minimum fatal dose (LD ₅₀) via oral administration is more than 60g/kg of extract. <Formulations> Preparation of pharmaceutical composition and food composition

<Formulation Example 1> Preparation of capsules 1 OOmg of acanthopanax extract, 1 OOmg of cornstarch, 1 OOmg of lactose and 2mg of magnesium stearate were completely mixed, followed by filling into a hard gelatin capsule, thereby forming a hard gelatin capsule formulation.

<Formulation Example 2> Preparation of injection Using l OOmg of acanthopanax extract, adequate amount f distilled water and adequate amount of pH regulator, acanthopanax extract was dissolved into distilled water for injection and regulated at the degree of about 7.5 pH, followed by filling in 2m/ample with distilled water and sterilizing, thereby forming an injection formulation. <Formulation Example 3> Preparation of health-drinkables Sub-ingredients such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%>), and water (75%) and acanthopanax extract were homogeneously compounded and instantly sterilized, followed by filling in the small receptacles of glass bottles, pet bottles, etc, thereby forming a health-drinkables formulation.

Industrial Applicability As aforementioned, the composition according to the present invention can provide a composition for preventing or treating dementia comprising acanthopanax extract as an effective ingredient. In the present invention, the composition for preventing or treating dementia can be embodied into a pharmaceutical composition for preventing or treating dementia or a food composition for preventing or treating composition. Although, hitherto, the present invention was described by way of embodiments, it is also evident that any various modifications can be made without departing from the scope of the present invention. According, the scope of the present invention should not be limited within the extent of the explained embodiments, and should depend not only on claims to be mentioned later, but also on things equivalent to this scope of the claims.

Claims

Claims 1. A composition for preventing or treating dementia comprising acanthopanax extract prepared by extracting acanthopanax using water, distilled water, alcohol, nucleic acid, ethyl acetate, acetone, chloroform, methylene chloride or mixtures thereof, as an effective ingredient.

2. The composition according to claim 1, comprising: a composition for preventing or treating dementia characterized by a pharmaceutical composition.

3. The composition according to claim 1, comprising: a composition for preventing or treating dementia characterized by a food composition.

4. The acanthopanax according to either of claim 1 or claim 3, comprising: a composition for preventing or treating dementia characterized by acanthopanax divaricatus or acanthopanax senticosus.

5. The acanthopanax extract according to either of claim 1 or claim 3, comprising: a composition for preventing or treating dementia characterized by that acanthopanax divaricatus or acanthopanax senticosus was extracted using distilled water or alcohol aqueous solution.

6. The acanthopanax extract according to either of claim 1 or claim 3, comprising: a composition for preventing or treating dementia characterized by that acanthopanax divaricatus or acanthopanax senticosus was extracted using methanol aqueous solution or ethanol solution.

7. The acanthopanax extract according to either of claim 1 or claim 3, comprising; a composition for preventing or treating dementia characterized by the selection of the group following (i), (ii), or (iii) below. (i) acanthopanax extract prepared by boiling-extracting acanthopanax divaricatus or acanthopanax senticosus with distilled water; (ii) acanthopanax extract prepared by stirring acanthopanax divaricatus or acanthopanax senticosus with ethanol aqueous solution; and (iii) acanthopanax extract prepared by stirring acanthopanax divaricatus or acanthopanax senticosus with methanol aqueous solution.

8. The dementia according to either of claim 1 or claim 3, comprising: a composition for preventing or treating dementia characterized by Alzheimer's disease.