KR100799884B1 - Composition for preventing or improving the ischemia damage compring an extract of epimedium koreanum - Google Patents

Abstract

The present invention relates to a composition for preventing or improving ischemic cerebrovascular disease, including trilobite leaf extract, and more particularly, to effectively prevent neuronal cell damage in the CA1 region of hippocampus known to be sensitive to cerebral ischemia, and to prevent side effects on the human body. It is possible to provide a composition for preventing or improving ischemic cerebrovascular disease, including harmless trigeminal vinegar extract.

Trilobite leaf, ischemic cerebrovascular disease, dementia, Alzheimer's disease, memory

Description

COMPOSITION FOR PREVENTING OR IMPROVING THE ISCHEMIA DAMAGE COMPRING AN EXTRACT OF EPIMEDIUM KOREANUM}

Figure 1 is a measure of lactate dehydrogenase (LDH) by applying a hypoxic environment to PC12 cultured cells to prevent cell death of trifoliate vinegar extract.

FIG. 2 shows the hippocampal tissues of the normal group (A and B) and the solvent (water) administration group (C and D) when subjected to ischemia-reperfusion, using an axiphot microscope.

Figure 3 is orally administered to the experimental animals at 50 mg / kg or 100 mg / kg daily from a week before the ischemia-reperfusion the trigeminal foliar extract to sacrifice the animals and stain the hippocampus tissue with cresyl violet (cresyl violet) Observed.

Figure 4 shows the survival rate of neurons by sacrificing the animals 4 days after the administration of 50 mg / kg or 100 mg / kg daily to a week before the ischemia-reperfusion the trigeminal leaf extract.

[Industrial use]

The present invention relates to a composition comprising tricotyledonous extract having a neuroprotective effect, and may be used as a food or pharmaceutical composition.

[Private Technology]

Of the total population released in 2005, the National Statistical Office recently made up 9.1% of the population aged 65 and over, and the proportion of young people aged 14 and younger is decreasing. The proportion of the population aged 65 and over is 7.2% in 2000. Has already entered an aging society. In addition, as of 2003, the average life expectancy of Korea is 77.5 years, and the elderly population is rapidly increasing.

Meanwhile, Korea's GNI per capita, announced by the National Statistical Office, was $ 14,162 in 2004, and is increasing by more than 10% annually. Accordingly, the number of deaths from infectious diseases, which are the major causes of death, has declined sharply in the past, and conversely, degenerative diseases such as cancer, cerebrovascular diseases, and diabetes have been steadily increasing. Cerebrovascular disease is the second largest cause of death in Korea and is caused by a decrease in blood flow to the brain.

Ischemic disease is a disease caused by delayed neuronal death in the hippocampus CA1 region due to a decrease in blood flow in the brain and lack of oxygen and glucose (Kirino, Brain Res ., 239 , pp57-). 69, 1982; Petito et al., Neurology , 37 , pp1281-1286, 1987; Pulsinelli et al., Ann. Neurol . , 11 , pp491-498, 1982). In addition, the ischemic brain disease is known to be caused by the lack of blood supply to the brain in pathological conditions such as stroke or heart attack (Nedergaard, Acta . Neurol . Scand . , 77 , pp81-101, 1988; White et. al., Neurology , 43 , pp 1656-1665, 1993).

The cause of delayed neuronal death in the hippocampal CA1 region caused by transient cerebral ischemia and reperfusion is caused by the influx of calcium into cells (Hara et al., Brain Res. Bull. , 29 , pp659-665, 1992; Nedergaard , Acta . Neurol . Scand . , 77 , pp81-101, 1988), known as free radical-related neuronal damage and glutamate-receptor mediated neuronal damage (Won et al., Neurosci . Lett . , 301) . , pp 139-142, 2001). That is, when cerebral ischemia occurs, depolarization of neurons occurs, and glutamate of synapses is released, thereby increasing the concentration of extracellular glutamate. In addition, the reversal of active transport caused by the reduction of Adenosine Triphosphate (ATP) accelerates the accumulation of extracellular glutamate. Accumulation of extracellular glutamate is characterized by N-methyl-D-aspartic acid (NMDA), α-amino-3-hydroxy-5-methyl-4 -isoxazole-propionic acid (AMPA), and metabotropic glutamate receptor (MTA). Induces a continuous activity of calcium, leading to an influx of calcium into the cells (Olney et al., Science , 244 , pp1360-1362, 1989). Intracellular influx of calcium triggers the activity of calcium-dependent proteases, lipases and modulators, and in turn leads to the generation of cytotoxic molecules, including free radicals, that break down cellular structures such as DNA and cell membranes, causing neuronal death. .

Recently, nerve cells have been reported to regenerate, but the prevention is more important than the treatment due to the histological characteristics of regeneration difficult. Once cerebrovascular disease, irreversible nerve cell damage is known to cause a decrease in motor performance, sexual performance, memory loss, etc., the importance of prevention is emphasized. Therefore, it can be said that it is important to prevent such cerebrovascular diseases by continuously ingesting substances that can protect the nerve cells of the brain.

Cerebrovascular disease is less effective for treatment, and efforts are underway to find active ingredients from natural-derived materials.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or improvement of ischemic cerebrovascular disease, including trichophytic vinegar extract, which has no toxicity to the human body and has a significant neuroprotective effect.

In addition, an object of the present invention is to provide a food composition for the prevention or improvement of ischemic cerebrovascular disease, including trilobite vinegar extract.

The present invention provides a composition for the prevention or improvement of ischemic cerebrovascular disease comprising tricotyledonous extract having an effect of nerve cell protection as an active ingredient in order to achieve the above object.

In another aspect, the present invention provides a pharmaceutical composition for preventing or improving ischemic cerebrovascular disease comprising tricotyledonous extract as an active ingredient having neuroprotective effect.

The present invention provides a food composition for preventing or ameliorating ischemic cerebrovascular disease, including tricotyledonous extract having an effect of protecting nerve cells as an active ingredient. Examples of the food composition of the present invention include food, food additives, or beverages.

Hereinafter, the present invention will be described in more detail.

Trilobite leaf ( Epimedium) of the present invention koreanum ) is a perennial herb and grows in the shade of mountain forests around 30cm in height. The leaves are regenerated and split three times, and three branches and nine leaves are named. The lobules are ovate, pointed at the end, and have hairy cogs on the edges. The flower is yellowish white and reddish purple, and it runs in the form of gunshot inflorescence. Trichophytium is used as a medicinal herb in tonic, gangjeong, diuretic, emphysema, forgetfulness, and vulgaris.

Tricuspid foliar extract of the present invention may be a dry powder of a solvent extract, a fraction extract or a liquid thereof.

Extraction solvent of the present invention is water; Lower alcohols having 1 to 4 carbon atoms such as methanol and ethanol; Polar solvents such as ethyl acetate; Nonpolar solvents of hexane and dichloromethane; Or a mixed solvent thereof. Preferred extraction solvents of the present invention are lower alcohols having 1 to 4 carbon atoms, such as 50 to 100% methanol or ethanol, and more preferably 70 to 100% methanol or aqueous methanol solution.

Tricuspid leaf extract of the present invention can be obtained as follows.

The trigeminal leaf vinegar extract of the present invention can be extracted using a solvent having a volume of about 1 to 15 times the dry weight, preferably about 2 to 7 times the dry weight, by slicing the leaves and root portions of the trigeminal vinegar harvested and dried. The solvent may be used for extraction for 1 to 12 hours, preferably about 1 to 5 hours, at an extraction temperature of 50 to 90 ° C. As the extraction method of the extract, any extraction method using a conventional solvent may be applied. For example, an extraction method such as dipping, ultrasonic extraction, and reflux extraction may be used.

In addition, the trigeminal leaf vinegar extract may further perform a conventional fractionation process. Examples of fractionation methods of extracts are described in, but not limited to, Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis, 3 ^rd Ed., Pp 6-7, 1998.

The obtained extract or fractions may be concentrated and lyophilized to obtain a powdery trichophytic herb extract from which moisture is completely removed.

The present invention provides a medicament or plant composition for the prevention or improvement of cerebral ischemic disease comprising the trigeminal leaf herb extract.

The ischemic cerebrovascular disease may be Alzheimer's disease, dementia or stroke. Trilobite leaf extract of the present invention anesthetize gerbils to induce cerebral ischemia by ligation of allergic arteries, and then staining nerve cells to measure their efficacy and exposing PC12 cells, which are adrenal derived neuronal cell lines to hypoxic environment, to be released from cells. In all experiments measuring the effect of lactate dehydrogenase concentration, it significantly inhibited neuronal cell death caused by cerebral ischemia and showed an excellent neuronal protective effect (Koo BS, Lee WC, Chung KH, Ko JH, Kim). CH.A water extract of Curcuma longa L. (Zingiberaceae) rescues PC12 cell death caused by pyrogallol or hypoxia / reoxygenation and attenuates hydrogen peroxide induced injury in PC12 cells.Life Sci. 2004 Sep 24; 75 (19): 2363-75) .

The composition for preventing or improving cerebral ischemic disease of the present invention comprises 0.001 to 99.99% by weight, preferably 0.1 to 90% by weight, based on the total weight of the composition.

The pharmaceutical composition or food composition comprising the tricotyledonous extract of the present invention may further include appropriate carriers, excipients and diluents commonly used in the preparation thereof.

The dietary supplement composition comprising the extract according to the present invention may be used in oral formulations, such as powders, granules, tablets, suspensions, emulsions, and syrups, respectively, according to conventional methods. Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose And methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solvents, emulsions, and serums.In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight, extent of disease, drug form, route of administration and duration, but may be appropriately selected by those skilled in the art. Preferably the extract of the present invention is 0.0001 to 1000mg / kg per day, in order to be more effective it is good to administer at 0.01 to 100mg / 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 composition of the present invention can be used as food. As used herein, the term "food" is intended to include all foods, food additives, and beverages.

Examples of the food to which the tricotyledonous extract of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods. In addition, the food in the present invention includes special nutritional products (e.g., prepared oils, infants, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasoned foods ( For example, soy sauce, miso, red pepper paste, mixed soy sauce), sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, and the like, and natural seasonings (eg, ramen soup). The food, beverage or food additives may be prepared by a conventional manufacturing method.

In addition, in the food composition aimed at the effect of preventing or improving cerebral ischemic disease, the amount of the extract may include 0.01 to 15% by weight of the total food weight, the beverage composition is 0.02 to 5 based on 100 ml g, preferably 0.3 to 1 g.

The beverage composition may include additional components in addition to the extract, for example, various flavors or natural carbohydrates. 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 described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention, in addition, the extracts of the present invention are pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. It may further contain.

In addition to the above, the extract of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. These components can be used independently or in combination. The proportion of such additives is not so critical, but may be selected in the range of 0 to 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, preferred embodiments of the present invention are described. However, the following examples are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the following examples.

Example  1: Preparation of Trichophyte Leaf Extract

After trituration of 100 g of dried trigeminal vinegar (Cheorwon, Gangwon-do) finely, 600 ml of 100% methanol was added thereto, followed by immersion at 74 ° C. for 5 hours. Methanol in the extract was completely removed using a vacuum rotary evaporator and concentrated, followed by complete removal of moisture for 48 hours using a lyophilizer to obtain Trichophyte leaf powder (8.74 g), and 1/10 of the extract obtained was 10 It was used for the experiment as a stock solution (98 mg extract / ml water) that was dissolved in ml of water.

Example  2: Trichophyllum Extract Cerebral ischemia  Nerve cell protective effect

In order to measure the prevention and improvement of neuronal cell death by cerebral ischemia, male Mongolian gerbil (Harlan, USA), weighing 65-75 g, was anesthetized and ligated into the common carotid artery to induce cerebral ischemia. Then, the method of confirming by neuronal staining was used. In addition, the efficacy of the Trichophyte herb extract of Example 1 was verified by two methods of inducing a hypoxic environment using an adrenal derived PC12 cell line and then measuring lactate dehydrogenase concentration.

2-1: Lactate Dehydrogenase (in vitro) of Trichophytium Echo Extract by Concentration Measurement Cerebral ischemia  Nerve cell protection efficacy measurement

Lactide dehydrogenase (LDH) is secreted from the cultured cells into extracellular culture to induce neuronal damage through hypoxic environment by injecting CoCl ₂ using PC12 cells. The concentration was measured. PC12 cells were cultured in a medium containing 7% fetal calf serum, 7% horse serum, 100 ug / ml streptomycin and 100 unit / ml penicillin at 37 ° C and 6% carbon dioxide. Cell culture medium was collected at 20 hours when the secretion was almost completed, and the result was shown in FIG. 1 by measuring with a microplate reader.

The tricotyledonous extract obtained in Example 1 was treated with the culture medium, 10, 50, 100, 500, 1000 and 5000 μM, respectively, and then treated with 150 μM CoCl 2 after 30 minutes. PC12 cells were cultured at 37 ° C. for 20 hours to obtain cell culture medium, and Beckman DU-640 Absorbance Spectrometer using lactate dehydrogenase concentration in the culture medium as Zhong Sheng Biotech standard. Was measured by enzyme dynamic method (Hou et al. J. Neurosci. Res., 74, pp123-133, 2003; Yamakawa et al. Neurol. Res., 23, pp522-530, 2001). .

As a result, as shown in FIG. 1, the treatment rate of the triglyceride vinegar decreased the concentration of lactate dehydrogenase in the group treated with the solvent only and then compared to the control group that induced the hypoxic environment. As a result, it was confirmed that the trigeminal leaf vinegar extract has a neuroprotective effect and an improvement effect.

2-2. Trichophytium Leaf Extract through Animal Experiment Cerebral ischemia  Nerve cell protection efficacy measurement

Animal experiments using gerbils confirmed the neuronal cell death inhibitory effect of Trichophytium foliar extract.

2-2-1. Breeding of Experimental Animals

40 male Mongolian gerbil ( Meriones unguiculatus, Halan, USA), weighing 65-75 g, were used. Experimental animals were subjected to a temperature of 23 ± 2 ° C. under constant light and dark cycles from 7 am to 7 pm. Breeding at a humidity of 55 ± 10%. Food and water were taken freely.

2-2-2. Evaluation of Neuroprotective Effect of Trigeminal Leaf Herb Extract in Brain Ischemia

The trigeminal leaf vinegar extract of Example 1 was dissolved in distilled water in experimental animals 0.5 ml orally administration (e.g., 50 mg or 100 mg / kg body weight) using an esophageal needle every day from one week before ischemia induction. The animals were subjected to general anesthesia with 3% isoflurane (isoflurane, Baxtor, USA) in a gas mixture of nitrogen and oxygen 7: 3, and the operation was performed while maintaining anesthesia with 2.5% isoflurane in the same gas. . Both hairs of the neck were cut and disinfected, and then incision was made to expose both warm arteries, ligation was performed for 5 minutes using an aneurysm clip (aneurysm clip, Staelting, USA), and then the clips were removed and reperfused. At this time, each experimental group confirmed the complete occlusion of the entire artery by observing the blood circulation of the central artery of retina using an ophthalmoscope. The body temperature was measured by inserting a rectal thermometer during induction of ischemia, and the body temperature was kept constant at a normal body temperature of 37 ± 0.3 ° C by using a heating pad which is automatically adjusted according to the temperature of the experimental animal.

The experimental group was anesthetized by intraperitoneal injection of thiopental sodium at 30 mg / kg body weight 4 days after induction of cerebral ischemia, and then at 4 ° C. containing 1,000 IU per 1,000 ml. Physiological saline was injected into the left ventricle and washed perfusion. After the perfusion wash, the animals were directly perfused with 4% paraformaldehyde (in 0.1 M phosphate buffer; PB, pH 7.4) at 4 ° C.

After perfusion fixation, the animals were opened by using a bone cutter to remove the brain, and then fixed in the same fixative for 6 hours. After fixation, the brain was settled in a 30% sucrose solution (in 0.1 M phosphate buffer) until it settled to the bottom, and the tissue was cut into a 30 μm thickness using a sliding microtome (Reichert-Jung, Germany). 6-well plate containing a storage solution (storing solution) was stored at 4 ℃ until use.

Of each tissue section, a tissue having a well-formed hippocampal formation was selected, and washed three times with 0.01 M PBS for 10 minutes to remove the preservative solution from the tissue. It was smeared onto gelatinized slides, dried sufficiently at 37 ° C., and then immersed in distilled water for a while, and then stained with a 2% cresyl violet acetate (Sigma, USA) solution for 1 minute. The tissue is washed thoroughly with running water to remove excess dye from the slides, briefly immersed in distilled water, and then dehydrated and excess cresyl through 50%, 70%, 80%, 90%, 95% and 100% solutions. Violet was washed. After confirming that the tissue was visible in the Nissle body (Nissle body), immersed in xylene (Junsei, Japan) and made transparent and then encapsulated in Canada Balsam (Kanto, Japan).

The control group was prepared in the same manner as above except for the administration of distilled water used to dissolve the trigeminal vinegar extract, except that the normal group was not administered the trigeminal vinegar extract and did not undergo ischemia-reperfusion surgery. It prepared in the same manner as above.

Each tissue was photographed using the Axiphot microscope (Carl Zeiss, Germany) with a digital camera (Axiocam, Cal Zeiss, Germany) at 25 times the total hippocampal area and 200 times the CA1 area. The group is shown in Figure 2, the experimental group administered with the three trichophytium extract is shown in Figure 3.

2 and 3, A and C are photographs of the entire hippocampal region, and B and D represent CA1 regions of the hippocampus. In the normal group, the control group, and trigeminal vinegar when 50 mg / kg and 100 mg / kg were administered, purple stained portions were shown in FIG. 4 by counting nerve cells using an image analyzer (Optimas 6.5, USA) program. . One-way ANOVA test was performed to verify the significance of each group, and data were expressed as mean and standard mean error. In particular, FIGS. 2 and 3 were taken with the most common portion of each group using an axiophot microscope (Carlises Zeiss, Germany).

2-2-3. Experiment result

As shown in Figure 1, when administration of the tricotyledonous extract of 50 to 100 mg / kg was observed that the production of lactate dehydrogenase in PC12 cells is small, the nerve cell protective It was found to be effective.

In addition, in FIG. 2, the normal group was able to observe the deep staining of nerve cells in the whole area of the hippocampus, whereas in the control group, cells stained by ischemia in the CA1 region of the hippocampus were hardly observed.

On the other hand, in the experimental group administered the triglobin vinegar extract of 50mg / kg and 100mg / kg of Figure 3 it can be confirmed that the cells in the whole area of the hippocampus stained dark, trigeminal vinegar significantly inhibited neuronal cell death was confirmed. .

In addition, even in the results of confirming the neuronal cell death according to the concentration of tricotyledonous extract after the ischemia-reperfusion of FIG. 4 by the neuronal cell count, the CA1 region cells of the hippocampus were observed about 11.3% compared to the normal group in the control group. The neuronal survival rate was higher in the group administered with Trifolium vinegar extract than the control group. Thirty-one percent of cells were observed when 50 mg / kg of Trichophyte herb extract was administered compared to the normal group, and as shown in FIGS. 3A and 3B, many cells were observed to die inside the hippocampus CA1 region (FIG. 3A). , B and 4). On the other hand, when administered at a dose of 100 mg / kg, 73% of cells were observed compared to the normal group, and similar numbers of cells were observed in all parts of the CA1 region as shown in FIGS. 3C and 3D (FIGS. 3C, D and 4). In addition, no side effects were observed upon administration at these doses.

Example  3. Acute Toxicity Test

Acute toxicity test was performed using 6-week-old specific pathogen-free (SPF) SD rats (Sprague-Dawley, Harlan, USA). Five animals of each group were orally administered to the trigeminal vinegar extract of the present invention at a dose of 50 mg / kg, 100 mg / kg, 200 mg / kg, and 400 mg / kg. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to observe abdominal organs and thoracic organ abnormalities.

As a result, no significant clinical symptoms or dead animals were noted in all animals treated with the test substance, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings. As a result, the trigeminal leaf herb extract of the present invention did not show toxic changes up to 400 mg / kg in rats, respectively, and the minimum lethal dose (LD ₅₀ ) was determined to be a safe substance of 400 mg / kg or more.

TABLE 1

Weight and Tissue Autopsy Findings of Various Organs by Concentration Following Administration

density Weight change Tissue autopsy findings Gastrointestinal tract 50 - - 100 - - 200 - - 400 - - brain 50 - - 100 - - 200 - - 400 - - kidney 50 - - 100 - - 200 - - 400 - - liver 50 - - 100 - - 200 - - 400 - -

TABLE 2

Changes in Blood Biochemistry and Hemocytosis According to Concentration of Trilobite Leaf

50 mg / kg 100 mg / kg 200 mg / kg 400 mg / kg Erythrocyte Morphology 8.8 X 10 ⁶ / mm ³ 8.5 X 10 ⁶ / mm ³ 8.9 X 10 ⁶ / mm ³ 8.2 X 10 ⁶ / mm ³ Leukocyte count change 11.5 X 10 ³ / mm ³ 14.2 X 10 ³ / mm ³ 15.3 X 10 ³ / mm ³ 13.7 X 10 ³ / mm ³ Behavioral change - - - - Erythrocyte Morphology - - - -

From the above experimental results, pathological findings were not found even when administered in different concentrations in animal experiments by inhibiting neuronal cell death by cerebral ischemia, and there was no change in weight of organs and no significant change in blood chemistry. The composition comprising the same may be usefully used as a dietary supplement for the prevention and improvement of cerebral ischemic disease.

Formulation example  One. Powder  Produce

Tricuspid leaf extract powder 20 mg

Lactose 100 mg

Talcuk 10 mg

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

Formulation example  2. Preparation of Tablets

Tricuspid leaf extract powder 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

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

Formulation example  3. Preparation of Capsule

Tricuspid leaf extract powder 10 mg

Crystalline Cellulose 3 mg

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. Liquid  Produce

Tricuspid leaf extract powder 20 mg

Isomerized sugar 10g

5 g of mannitol

Appropriate amount of purified water

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the mixture, and then the above ingredients are mixed, purified water is added to adjust the total amount to 100 ml, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation example  5. Manufacture of food

Tricuspid leaf extract powder 1000 mg

Proper amount of vitamin mixture

70 ㎍ 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 ㎍ of vitamin B12

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinamide 1.7 mg

Folic acid 50 ㎍

Calcium pantothenate 0.5 mg

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

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above components are mixed according to a conventional food production method. Granules may be prepared and used to prepare food compositions according to conventional methods.

As described above, the trigeminal leaf vinegar extract of the present invention not only effectively prevents damage to nerve cells in the CA1 region of the hippocampus known to be sensitive to adrenal-derived cell lines and cerebral ischemia, but also inhibits neuronal cell death caused by cerebral ischemia and has side effects on the human body. It can be usefully used as a dietary supplement for the prevention and improvement of harmless cerebral ischemic disease that does not occur.

Claims (7)

A pharmaceutical composition for preventing or treating stroke through nerve cell protection, containing trigeminal leaf vinegar (Epimedium Koreanum) methanol extract as an active ingredient. The pharmaceutical composition for preventing or treating stroke according to claim 1, which contains 0.001 to 99.99% by weight of the tricotyledonous methanol extract. delete delete delete Claim 3 or claim 3 comprising an extract of Trimedium Korean herb (Epimedium Koreanum) methanol as an active ingredient, a food composition for preventing or improving stroke. The food composition of claim 6, wherein the food composition is a food, a food additive, or a beverage.

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