KR20110134641A - Pharmaseutical compositions for prevention or treatment of cerebrovascular disease, or for improving impairments, containing the extracts of curcuma longae radix, phellinus linteus and scutellariae radix as an active ingredient - Google Patents

Abstract

PURPOSE: A composition containing Curcuma Longae Radix, Phellinus Linteus, and Scutellariae Radix is provided to suppress Ca^2+ and reactive oxygen generation. CONSTITUTION: A pharmaceutical composition for preventing or treating cerebrovascular diseases contains extract of Curcuma Longae Radix, Phellinus Linteus, and Scutellariae Radix as an active ingredient. The extract is isolated using water, alcohol, or mixture thereof. The alcohol is C1-C4 low alcohol. A neutraceutical food for preventing and treating cerebrovascular diseases contains Curcuma Longae Radix, Phellinus Linteus, and Scutellariae Radix as an active ingredient.

Description

Pharmaceutical compositions for prevention or treatment of cerebrovascular disease, or for improving impairments, containing the extracts of Curcuma Longae Radix , Phellinus Linteus and Scutellariae Radix as an active ingredient}

The present invention relates to a pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage containing a plant extract as an active ingredient.

Cerebral apoplexy, or cerebral infarction, is the main cause of thromboembolism of the cerebral arteries due to vascular atherosclerosis or from cardiac embolism due to embolism and heart disease. Cerebral infarction, which is caused by clogged cerebrovascular vessels, is further divided into cerebral thrombosis and brain embolism. Cerebral thrombosis is caused by arteriosclerosis due to hypertension, diabetes, hyperlipidemia, etc., and the walls of the arteries become thick or hard. As a result, blood vessels are narrowed and the inner wall of the blood vessels are not smooth, so blood is entangled and clogged, and blood supply is significantly reduced or stopped, resulting in a lack of oxygen and nutrient supply to brain cells, resulting in impaired brain function. Cerebral embolism causes abnormal blood flow in the heart due to diseases such as heart valve or atrial fibrillation, which causes a part of the blood to stop partially in the heart, and thus clots, resulting in blood clots. Cerebral infarction develops.

Vascular dementia is an impairment of acquired irreversible cognitive function caused by brain injury caused by cerebrovascular disease. Dementia caused by stroke and cerebral infarction account for about one-third of all dementia, with the highest frequency after Alzheimer's. As in general stroke and cerebral infarction, patients with vascular risk factors such as old age and smoking are known to have large or small vessel occlusion of the brain, which causes vascular dementia as the cortical or subcortical neuronal fibers are destroyed. have.

Vascular dementia is caused by multiple-infarct dementia, subcortical ischemic vascular dementia, and a single lesion due to occlusion, depending on the underlying vascular pathology or the location of the lesion causing the dementia. dementia). Among these, dementia due to a single lesion due to occlusion means dementia due to a single lesion occurring in the caudate nucleus, the mesial frontal lobe, the genu of internal capsule, and the like. White matter hyperintensity (leukoaraiosis), which is caused by a ruptured cerebral infarction or an incomplete infarction caused by small vessel disease, mainly damages the prefrontal-subcortical circuit. As a result, subcorticaldysfunctio, such as bradyphrenia and apraxia, may appear rather than symptoms of traditional dementia such as memory disorders.

Alzheimer's disease (AD) is characterized by neuronal loss and extracellular senile plaques, whose major component is amyloid-beta (Aβ), a 39-43 amino acid peptide derived from amyloid precursor protein. senile plaque). In vitro and in vivo studies have shown that Aβ or Aβ peptide fragments have toxic effects, suggesting that Aβ plays an important role in the development of AD (Butterfield et al. al . , Free Radical Biology and Medicine , 2002, 32: 1050-1060; ButterfIeld et al ., Free Radical Biology and Medicine , 2007, 43: 658-677). In culture, Aβ directly induces neuronal death and renders the neurons vulnerable to excitatory toxicity and oxidative damage. N-methyl-D-aspartate receptor (NMDA) receptors act as selective substrates of Aβ binding or mediators of Aβ-induced glutamate excitatory toxicity. NMDA receptors are highly transmissive in particular ligand in Ca ^{2 +} - gate / overvoltage - is sensitive cation channels. Extensive increase in [Ca ^{2 +]} _i will lead directly to cell dysfunction, over-excitement or death. Thus, the neurotoxic effect of Aβ is (5R.10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo [a, d] cyclohepten-5,10, a non-competitive NMDA receptor antagonist. Reduced by imine maleate (MK-801) [5R.10S]-(+)-5-methyl-10,11-dihydro-5H-dibenzo (a, b) cyclohepten-5,10-imine maleate] as will be demonstrated by the report, Ca ^{+ 2} influx through NMDA receptors by exposure Aβ plays a crucial role in the neurotoxicity induced Aβ-. The formation of reactive oxygen species (ROS) is also believed to be involved in the development of degenerative brain diseases. Some evidence supports the involvement of oxidative stress as an active factor in Αβ-mediated neuropathy by triggering or facilitating neurodegeneration through a wide range of molecular shapes that interfere with neuronal homeostasis. However, the clinical benefits of NMDA receptor antagonists and direct blockers of neuronal channels are controversial, as they lack the identifiable efficacy or have serious side effects.

Most degenerative brain diseases involve dementia, especially cognitive and memory disorders (memory decline). Therefore, a desirable therapeutic agent for degenerative brain diseases including dementia should delay brain cell destruction and aging to protect brain cells and restore cognitive function. Drugs developed to date include antioxidants such as vitamin E and selegiline, which inhibit the destruction of brain cells by free radicals, and acetylcholine such as tacrine, aricept and exelon Acetylcholinesterase-inhibiting drugs. However, such drugs have been reported to cause significant side effects, the disadvantage of which is not good efficacy.

Curcuma Longae is a perennial plant classified as the Curcuma of the Zingiberaceae. It is cultivated in India, China, Southeast Asia, and is used as an edible, medicinal and natural dye. Recently, various papers and media have reported that curcuminoids and curcumin have various physiological effects such as antioxidant, anti-cancer, anti-mutant, anti-inflammatory, dementia prevention, liver function protection and cholesterol reduction (Guo LY et. al .., 2008, Archives Pharmacal Research , 31: 490-496; Adaramoye OA et al ., 2002, Biokemistri 12: 127-135).

The situation is Phellinus sp ), mud mushrooms are known around the world, such as horse mud mushrooms ( P. igniarius ), clay mushrooms ( P. robustus ), larch mud mushrooms ( P. pine ). The situation was excellent in anti-immune function, gastrointestinal function activation, detoxification effect and 96.7% anti-cancer effect in animal experiment (Ajith TA et al ., 2002 Journal of Ethnopharmacology , 2002, 81: 387-391; Sliva et al . , 2008, British Journal of Cancer , 98: 1348-1356). Compared to other mud mushrooms, the effect is excellent. In addition, the situation has been proved in clinical trials, industrial application research is actively being conducted, and due to the availability of drugs, a large amount is distributed from China, India, and the like at a high price.

Golden ( Scutellariae Radix is a kind of herbal medicine that peels off the bark of dried Scutellaria baicalensis Gergi and dried roots. In general, Chinese medicine describes the inflammatory disease as a heat or fire, and the drug that has the action of treating it has a heat-fever metabolism effect by lowering the heat. In the past, it has been used as a clearing medicine, golden, yellow lotus, yellow white, rhubarb, gypsum, etc. Among them, the most representative gold is very hard in nature, so it is widely used in intestinal and lung recessive diseases. come. Thus, it was mainly applied to diseases such as fever, seawater, febrile diarrhea, jaundice, and hemostasis caused by external fraud invading the human body. Pharmacologically, baicalin, baicalein, and wogonin have been isolated, and branch offices, fever, anti-inflammatory and anticancer activity have been reported (Bensky D et. al ., 1992, Chinese Herbal Medicine Materia Medica . Eastland Press , pp . 107-109; Huang KC, 1999, The Pharmacology of Chinese Herbs , pp. 385).

Thus, the present inventors have self-cell death by beta amyloid in the brain cell culture are of, turmeric, conditions and Gold extract was efforts to develop natural material with a cerebrovascular disease preventing and treating effect from rat fetuses, Ca ^{2 +} generation and activity It is effective in preventing or treating cerebrovascular disease, or improving memory damage, and the pharmaceutical composition and health functional food by confirming that it inhibits oxygen production and effectively inhibits memory damage in mice inducing neurotoxicity by beta-amiroid. The present invention has been completed by confirming that it can be usefully used as a component.

The object of the present invention is Curcuma Longae Radix), situation ( Phellinus Linteus ) and Golden ( Scutellariae) Radix) to provide a pharmaceutical composition and health functional food for preventing or treating cerebrovascular disease or improving memory damage containing the extract as an active ingredient.

In order to achieve the above object, the present invention is curcuma Longae Radix), situation ( Phellinus Linteus ) and Golden ( Scutellariae) Provided is a pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage, containing the Radix extract as an active ingredient.

The present invention also provides a health functional food for preventing or improving cerebrovascular disease or improving memory damage, which contains turmeric, wheat and golden extract as active ingredients.

Curcuma of the present invention Longae Radix), situation ( Phellinus Linteus ) and Golden ( Scutellariae) Radix) extract inhibited autologous cell death, Ca ²⁺ production and free radical production by beta-amiroid of brain neurons cultured from rat fetuses, and effectively inhibited memory damage in beta-amiroid-induced neurotoxicity. It may be usefully used as an active ingredient in a composition for preventing or treating cerebrovascular disease, or a pharmaceutical composition for improving memory damage and a dietary supplement.

1 is Curcuma according to the present invention Longae Radix), Situations ( Phellinus Linteus ) and Golden ( Scutellariae) Radix) is a graph showing the inhibitory effect of Aβ (amyloid-beta) (25-35) -induced neuronal cell death.
Figure 2 is a graph showing the inhibitory effect of Aβ (25-35) -induced autologous apoptosis of turmeric, circumstance and golden extract according to the present invention.
Figure 3 is a graph showing the inhibitory effect of Aβ (25-35) -induced Ca ^{2 +} increase of turmeric, turmeric and golden extract according to the present invention.
Figure 4 is a graph showing the inhibitory effect of Aβ (25-35) -induced reactive oxygen species of turmeric, turmeric and golden extract according to the present invention.
Figure 5 is a graph showing the inhibitory effect of Aβ (25-35) -induced memory damage of turmeric, turmeric and golden extract according to the present invention.

Hereinafter, the present invention will be described in detail.

The present invention is curcuma Longae Radix), situation ( Phellinus Linteus ) and Golden ( Scutellariae) Radix) provides a pharmaceutical composition for the prevention or treatment of cerebrovascular disease or improvement of memory damage containing the extract (hereinafter referred to as HS0608) as an active ingredient.

Turmeric, situation and gold can be used without limitation, such as cultivated or commercially available, turmeric, situation and gold is preferably used in a weight ratio of 1: 1: 1, but is not limited thereto.

The HS0608 is preferably manufactured by a manufacturing method including the following steps, but is not limited thereto.

1) mixing turmeric, situation and gold;

2) extracting by adding an extraction solvent to the mixture of step 1);

3) cooling the extract of step 2) and then filtering; And

4) The filtered extract of step 3) may be prepared by sequentially condensing reflux and drying.

The extraction solvent is preferably water, alcohol or a mixture thereof. As the alcohol, C ₁ to C ₄ lower alcohols are preferably used, and as lower alcohols, ethanol or methanol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but is not limited thereto. The extraction solvent is preferably extracted by adding 5-15 times to the mixture of dried, turmeric and gold, and more preferably by adding 10 times. The extraction temperature is preferably 40 to 100 ° C, more preferably 60 to 80 ° C, but is not limited thereto. In addition, the extraction time is preferably 4 to 24 hours, more preferably 8 to 15 hours, but is not limited thereto. In addition, the number of extraction is preferably 1 to 5 times, it is more preferable to extract three times, but is not limited thereto.

In the above method, the drying of step 4) is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or lyophilization.

The cerebrovascular disease is preferably any one selected from the group consisting of cerebral infarction, cerebral fever, stroke, dementia, Alzheimer's disease, Huntington's disease, pick and Creutzfeld-Jakob disease, but not limited thereto. Do not.

In a specific embodiment of the present invention, brain neurons were isolated from the fetuses of rats and cultured and tested for efficacy of neurons. As a result, HS0608 suppresses the generation of my self apoptosis cells by Aβ (apoptotic cell death) (see Fig. 2), Ca ^{2 +} concentrations increase (see Fig. 3), the intracellular reactive oxygen species (ROS) (see Fig. 4) It was. Therefore, it was confirmed that HS0608 of the present invention inhibits neuronal cell death by Aβ, which is known to be a cause of dementia.

In addition, the present inventors conducted a passive avoidance test to confirm the inhibitory effect of HS0608 memory damage. As a result, the group to which HS0608 was administered showed suppression of memory damage in a dose-dependent manner (see FIG. 5).

Therefore, HS0608 of the present invention can be used as an active ingredient for the prevention or treatment of cerebrovascular diseases, or for improving the pharmaceutical composition for memory damage.

The composition containing HS0608 of the present invention is preferably included in 0.1 to 50% by weight based on the total weight, but is not limited thereto.

The compositions of the present invention can be administered orally or parenterally during clinical administration and can be used in the form of general pharmaceutical formulations. That is, the composition of the present invention may be administered in various oral and parenteral dosage forms in actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Is prepared using. Solid preparations for oral administration include tablets, pills, powders, granules and capsules, and the like, which may be used in the pharmaceutical composition of the present invention at least one excipient such as starch, calcium carbonate, sucrose, lactose And gelatin etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium, styrate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, 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. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as oliveyl, 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, glycerol, gelatin and the like can be used. The pharmaceutical composition of the present invention may be via subcutaneous injection, intravenous injection or intramuscular injection during parenteral administration.

Dosage units may contain, for example, 1, 2, 3 or 4 times the individual dosage or may contain 1/2, 1/3 or 1/4 times. The individual dosage preferably contains an amount in which the effective drug is administered at one time, which usually corresponds to all, 1/2, 1/3 or 1/4 times the daily dose. The effective dose of the composition of the present invention is 0.0001 to 10 g / kg, preferably 0.0001 g to 5 g / kg, may be administered 1 to 6 times a day.

The composition of the present invention can be used alone or in combination with methods using surgery, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of cerebrovascular diseases.

The present invention also provides a health functional food for preventing or improving cerebrovascular disease or improving memory damage, which contains turmeric, wheat and golden extract as active ingredients.

In a specific embodiment of the present invention, the present inventors have tested the efficacy of HS0608 after inducing toxicity to neurons in order to confirm the prevention and treatment effect of cerebrovascular disease of HS0608, HS0608 is intracellular autologous cell death by Aβ It inhibited the production of (apoptotic cell death), Ca ^{+ 2} concentration increases, the intracellular reactive oxygen species (ROS). Groups administered HS0608 in neurotoxic induced mice showed dose-dependent inhibition of memory damage.

Thus, the present invention can be used for preventing or improving cerebrovascular diseases containing turmeric, wheat and golden extract as an active ingredient, or health functional food for improving memory damage.

The health functional food of the present invention may be added as it is or used with other food or food ingredients, and may be appropriately used according to conventional methods. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). Generally, in the preparation of food or beverages, the composition of the present invention is added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight based on the raw materials. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

There is no particular limitation on the kind of food. Examples of the food to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, etc., includes all of the health food in the conventional sense.

The health beverage composition of the present invention may contain various flavors, natural carbohydrates, and the like as additional components, as in a conventional beverage. Natural carbohydrates described above are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin, cyclodextrin, sugar alcohols such as xylitol, sorbitol, erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, and synthetic sweetening agents such as saccharin and aspartame can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, HS0608 of the present invention can be used in various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents and the like used in beverages. In addition, HS0608 of the present invention may contain pulp for producing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is usually selected in the range of 0.02 to 1 parts by weight per 100 parts by weight of the composition of the present invention.

HS0608 of the present invention has little toxicity and side effects, so can be used with confidence even for long-term administration for the purpose of prevention.

Hereinafter, the present invention will be described in detail by Examples, Experimental Examples and Preparation Examples. However, the following Examples, Experimental Examples, and Production Examples specifically illustrate the present invention, and the contents of the present invention are not limited to Examples, Experimental Examples, and Production Examples.

< Example  1> curcuma ( Curcuma Longae Radix ), situation( Phellinus Linteus ) And golden ( Scutellariae Radix Manufacture of extract

<1-1> Preparation of Ethanol Extract

Dokbok, Situation and Gold were purchased from Gyeongdong Herbal Medicine Market in Seoul and used in small pieces. 100 g each of virulence, circumstance and gold were combined and extracted three times at room temperature in a reflux condenser with 3 L of 80% ethanol for 3 hours. The solution was filtered through Whatman No. 1 paper and dried to finally give 64 g of ethanol extract (HS0608). The final turmeric, turmeric and golden mixed extract (HS0608) were dissolved in dimethylsulfoxide (DMSO) at a concentration of 50 mg / ml and stored at -20 ° C. In experiments, the final concentration of DMSO was diluted to 0.1% or less in experimental buffer.

<1-2> Preparation of Water Extract

Extracted in the same manner as the extraction method of Example <1-1>, except that water was used instead of 80% ethanol as the extraction solvent to obtain a powder (yield 35.5%).

<1-3> Preparation of 100% Ethanol Extract

Except for using 100% ethanol instead of 80% ethanol as the extraction solvent was extracted in the same manner as the extraction method of Example <1-1> to obtain a powder (yield 26.4%).

<1-4> Preparation of 100 Methanol Extracts

Extracted in the same manner as the extraction method of Example <1-1>, except that 100 methanol was used instead of 80% ethanol as an extraction solvent to obtain a powder (yield 33.1%).

< Experimental Example  One > Culture of Cerebral Cortical Neurons

Primary cortical neurons were prepared by known methods from Sprague-Dawley rat fetuses of 15 to 16 days of age (Ban JY et. al ., Life science , 2006, 79: 2251-2259).

< Experimental Example  2 > HS0608 Of neuronal cell death inhibition

The present inventors performed the following experiment to confirm the effect of the turmeric, circumferential and gold mixed extract (HS0608) obtained in Example 1 on neuronal cell death.

Cortical neurons prepared in Experimental Example 1 were used in the experiment 3-4 days after the culture. None of the neurons were treated, or HS0608 obtained in Example 1 was treated with 0.1, 1, 10 and 50 mg / ml. After 15 minutes of treatment, 10 μM Aβ (25-35) (25-35) dissolved in serum-free Dulbecco's modified Eagle's medium (DMEM) to induce neuronal cytotoxicity (Bachem, Switzerland) ) Was incubated for 36 hours at 37 ℃ (Aβ treatment group and Aβ + HS0608 treatment group). Thereafter, 3- (4,5-dimethylthiazol-2-yl-2,5, diphenyltetrazolium bromide) [3- (4,5-dimethylthiazol-2-yl-2,5-diplenyltetrazolium by a known method bromide; MTT)] analysis was performed (Ban JY et. al ., 2006). At this time, when the Aβ-treated cells were used as a control group, the absorbances of the Aβ-only treatment group and the HS0608 treatment group were expressed as% in the graphs of FIGS. 1 and 1.

As shown in FIG. 1 and Table 1, in the 10 μM Aβ (25-35) alone group, the absorbance was reduced to 70.6% compared to the control (100%). This means that about 39% of neurons were killed by Aβ. By contrast, the absorbance of the HS0608 treatment group was 73.5% for 0.1 μg / ml treatment, 81.4% for 1 μg / ml treatment, 83.7% for 10 μg / ml treatment and 95.1% for 50 μg / ml treatment, respectively. Cell death was reduced in proportion to HS0608 concentration. In subsequent experiments, only the ethanol aqueous extract with the best cell death inhibition effect in MTT assay was tested.

Ethanol Aqueous Extract Water extract Ethanol extract Methanol extract  Control group 100 100 100 100  Aβ 71 72 72 72  Aβ + HS0608 0.1 74 73 72 73  Aβ + HS0608 1 81 74 75 78  Aβ + HS0608 10 84 77 76 79  Aβ + HS0608 50 95 79 81 82

< Experimental Example  3> HS0608 Neuronal Autologous cell death  Inhibitory effect measurement

The present inventors performed Hoechst 33342 staining by a known method on neurons treated in the same manner as Experimental Example 2 in order to prove that apoptosis by Aβ is autologous cell death (apoptosis) (Ban JY et. al ., 2006). The results are shown in FIG.

As shown in Figure 2, the control group caused 14.8% autologous cell death, whereas the Aβ-treated group caused 39.2% autologous cell death, and in the HS0608-treated group, 22.9% and 10 µg / ml at 1 µg / ml treatment. It was confirmed that autologous cell death occurred in 20.3% when treated with ㎖ and 13.4% when treated with 50 μg / ml and autologous cell death was reduced by HS0608.

< Experimental Example  4> HS0608 Inhibition of Calcium Increase in Cells

In order to confirm whether HS0608 of the present invention induces biochemical changes in cells, fluorine 4-AM (fluo-4 AM) (Molecular), which is a fluorescent staining agent, is used on neurons 3-4 days of culture prepared in Experiment 1 Probes Inc. USA) was added for 40 minutes to absorb cells, washed and treated nothing, or HS0608 obtained in Example 1 above was treated with 1, 10 and 50 μg / ml. After 15 minutes of treatment, immediately after treatment with Aβ (25-35) dissolved in 10 μM of serum-free DMEM in the neurons, changes in intracellular calcium concentration by Aβ (25-35) were determined by calcium-sensitive fluorescence. Fluorescent fluorescence is increased by the dye Fluo-4 AM (Molecular Probes, USA). Laser Scanning Conpocal microscope (LSM 510, Carl Zeiss, Germany) [488 nm Argon laser and 515 nm long. Pass radiation filter] was measured by intracellular calcium changes by a known method (Ban JY et. al ., 2006; Lee SB et al ., Korean Journal of Medicinal Crop Science , 2007, 15: 444-450). The measurement results are shown in FIG. 3.

As shown in FIG. 3, when Aβ was added to fluorine-4 AM-treated neurons, the intracellular calcium concentration ([Ca ^{2 +} ] _i ) increased and the fluorescence intensity started to increase when applied for 5 minutes. After increasing to the maximum value up to 153, the maximum value gradually decreased. On the other hand, if a pre-processing the HS0608 50 ㎍ / ㎖, it was confirmed that the increase in [Ca ^{2 +]} _i induced by Aβ is completely suppressed throughout the measurement period.

< Experimental Example  5> HS0608 Free radicals in ROS ) Measurement of production inhibitory effect

The present inventors treated the neuron prepared in Example 1 for 36 hours in the same manner as in Experiment 1 to confirm the inhibitory effect of HS0608 generation of neurons in neurons H ₂ DCF-DA (Molecular Probes) Inc., USA) for 10 minutes into the cells and using H ₂ DCF-DA by a known method in a laser scanning conpocal microscope MRC1021ES Bio-red (UK) [488 nm excitation and 515 nm emission filters]. ROS was measured as a change in fluorescence intensity by (Kim JY). et al ., Korean Journal of Medicinal Crop Science 2008, 16: 409-415, Lee SB et al ., 2007). The results are shown in FIG. 4.

As shown in FIG. 4, the fluorescence intensity was 242.8 due to the generation of reactive oxygen by Aβ, which is about 2.5 times higher than that of 47.6 of control cells. In comparison, HS0608 decreased fluorescence intensities to 138.7, 114.2 and 47.5, respectively, at 1, 10 and 50 μg / ml.

< Experimental Example  6> Manual Evasion Test Step-through Measurement of memory and learning by

Animals were used with ICR mice (BioLink Co. South Korea), HS0608 (25, 50 and 100 mg / kg po) orally administered on the first day of the experiment (day 1) after a period of adaptation after purchase, 30 needles of 27 gauge (gage) after minutes (Hamilton, USA) using a syringe equipped with a micro (microsylinge) was slowly injected with Aβ (25-35) 15㎕ 15 nmol of the ventricle (Maurice T et al ., Brain Reserch , 1996, 706: 181-193). Thereafter, HS0608 at 25, 50 and 100 mg / kg po was orally administered once daily for 1 week. HS0608 was orally administered on day 7, and 30 minutes later the mice were mastered in one trail step through passive-task. The passive evacuation box is a guillotine door divided into two sections: the grid floor and the impact generator. Place the mouse at the starting point in dark conditions, turn on the light after 50 seconds, open the door, and when the mouse enters the dark box, the door will close and electric stimulation (0.3 mA, 2 seconds) will be applied to the animal's dark box. It reminded me of my electric shock experience. After 24 hours, the animals were placed in bright boxes, and the same experiment did not move the animals that remembered the shock of the previous day to the dark box. The settling time of animals in bright boxes was measured and used as an indicator of memory formation. The results are shown in FIG. 5.

As shown in FIG. 5, the group not administered 15 nmol Aβ (25-35) moved to the dark place at 274 seconds, whereas the group administered with Aβ (25-35) moved to the dark room at 62 seconds. The results suggest that no memory was formed. On the other hand, the group receiving HS0608 25, 50, and 100 mg / kg daily for 1 week increased the time to enter the dark depending on the dose, so that the 25 mg / kg group received 165.4 seconds and the 50 mg / kg group received 222. Second, 100 mg / kg administration group was 279 seconds, all HS0608 administration group significantly increased, it can be seen that inhibits memory damage by Aβ (25-35).

< Manufacturing example  1>: Preparation of pharmaceutical preparation

<1-1> Preparation of Powder

HS0608 300 mg

Lactose 100 mg

Talc 10 mg

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

<1-2> Preparation of Tablet

HS0608 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components and tableting according to the conventional manufacturing method of tablets

To prepare.

&Lt; 1-3 > Preparation of capsules

HS0608 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

<1-4> Preparation of Injection

HS0608 50 mg

Appropriate sterile distilled water for injection

pH adjuster

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

<1-5> Preparation of Liquid

HS0608 1,000 mg

20 g of sugar

20 g per isomerization

Lemon flavor

Purified water was added to adjust the total volume to 1000 ml. The above components were mixed according to a conventional method for preparing a liquid, and then filled into a brown bottle and sterilized to prepare a liquid.

<Manufacture 2>: Preparation of health food

HS0608 1,000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixture is mixed with a composition suitable for a health food in a preferred embodiment, but the compounding ratio may be arbitrarily modified. The granules may be prepared and used for preparing a health food composition according to a conventional method.

< Manufacturing example  3>: Manufacture of healthy drinks

HS0608 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with a conventional method for preparing healthy beverages, and then stirring and heating 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 a healthy beverage composition of the invention.

Claims (10)

Curcuma Longae Radix), situation ( Phellinus Linteus ) and Golden ( Scutellariae) Radix) A pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage containing the extract as an active ingredient.

[Claim 2] The pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage of claim 1, wherein the extract contains turmeric, wheat and gold in a weight ratio of 1: 1: 1.
The pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage according to claim 1, wherein the extract is extracted with water, alcohol or a mixture thereof as a solvent.
The pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage according to claim 3, wherein the alcohol is C ₁ to C ₄ lower alcohols.
The pharmaceutical composition for preventing or treating cerebrovascular disease or improving memory damage according to claim 4, wherein the lower alcohol is ethanol or methanol.
6. The method of claim 1 or 5, wherein the cerebrovascular disease is from a group consisting of cerebral infarction, cerebral fever, stroke, dementia, Alzheimer's disease, Huntington's disease, pick and Creutz feld-Jakob disease. A pharmaceutical composition for preventing or treating cerebrovascular disease, or improving memory damage, characterized in that any one selected.
Health functional foods for the prevention or improvement of cerebrovascular disease or improvement of memory damage, containing turmeric, wheat and golden extract as active ingredients.
8. The dietary supplement for preventing or improving cerebrovascular disease or improving memory damage according to claim 7, wherein the extract contains turmeric, wheat and gold in a weight ratio of 1: 1: 1.
The health functional food for preventing or improving cerebrovascular disease, or improving memory damage, characterized in that the extract is extracted with water, ethanol or a mixture thereof as a solvent.
The method of claim 7, wherein the cerebrovascular disease is selected from the group consisting of cerebral infarction, cerebral fever, stroke, dementia, Alzheimer's disease, Huntington's disease, pick and Creutzfeld-Jakob disease. Health functional foods for preventing and improving cerebrovascular disease, or improving memory damage, characterized in that any one.

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