KR100669980B1 - Composition comprising an extract of Aurantii Immatri Pericarpium for the prevention and treatment of ischemic diseases and degenerative brain diseases - Google Patents

Abstract

The present invention relates to a composition containing the extract of Aurantii Immatri Pericarpium having the ability to improve cell survival under hypoxic conditions, and more particularly, the extract inhibits apoptosis in ischemic conditions and ischemic diseases and degenerative. It can be used in pharmaceutical compositions and health functional foods for the prevention and treatment of brain diseases.

Cheongpi, Apoptosis, Functional Food, Ischemic Disease, Degenerative Brain Disease

Description

Composition comprising an extract of Aurantii Immatri Pericarpium for the prevention and treatment of ischemic diseases and degenerative brain diseases}             

Figure 1a is a test performed using trypan blue dye exclusion (trypan blue dye exclusion) for the experimental group and the control group without the extract added to the different concentrations in the culture medium under hypoxic conditions,

Figure 1b is a diagram carried out using the trypan blue staining method for the experimental group and the control group without the extract was added to the extract of the different concentration in the culture medium under normal oxygen conditions,

Figure 2a is a diagram comparing the cell survival results of the cell line without the extract and the cell line added under hypoxic and normal oxygen conditions,

Figure 2b is a diagram showing the results of DNA fragmentation analysis of the cell line without the extract of the extract in culture medium under hypoxic conditions,

Figure 2c is a diagram showing the results of DNA fragmentation analysis of the cell line with the extract of the extract in culture medium under hypoxic conditions,

Figure 3a is a diagram comparing the results of the TTC (triphenyl tetrazolium chloride) staining of the cerebral infarction model and the myocardial infarction model not injected with dermal extract,

Figure 3b is a diagram showing the TTC staining results of the cerebral infarction model not injected with dermal extract,

Figure 3c is a diagram showing the results of TTC staining cerebral infarction model injected extract extract.

The present invention relates to extracts of the green skin having the therapeutic and prophylactic effect of ischemic diseases and degenerative brain diseases by inhibiting apoptosis.

Cerebral infarction and myocardial infarction, which are typical ischemic diseases, are blocked by atherosclerosis in which blood vessels are narrowed due to hypertension, hyperlipidemia, diabetes, and smoking. It is caused by necrosis of surrounding tissues.

Cardiovascular disease accounts for 30% of all mortality in the world, leading the mortality rate, and 75% of these are caused by ischemic diseases such as myocardial infarction and cerebral infarction. It is coming true. The methods of reducing the mortality caused by myocardial infarction and cerebral infarction include the treatment of hypertension and hyperlipidemia to prevent the occlusion of blood vessels and the reduction of necrosis of surrounding tissues during the occlusion.

The best way to reduce the necrotic area is to perforate the blocked blood vessels by thrombolytics, etc. in the earliest time, but re-perfusion the blood vessels, but within 3-6 hours of clogging the blood vessels as the heart and brain gain energy by breathing. The tissues are necrotic and subsequent reperfusion cannot be expected. However, it is practically difficult to arrive at the hospital within 3-6 hours of occlusion and undergo reperfusion after treatment, and the heart and brain are not regenerated once damaged. Therefore, if the cell viability in the ischemic state can be improved until reperfusion in the hospital can increase the therapeutic effect. At this time, the cause of tissue necrosis is apoptosis (Crow MT et al., Circ.Res ., 95 , pp957-970, 2004; Friedlander RM et al., N. Engl. J. Med. , 348 , pp1365- 1375, 2003), thereby inhibiting apoptosis, thereby improving cell viability.

In addition, in the case of surgery to stop the heart, if the amount of oxygen required more than the amount of oxygen supplied by the cardiopulmonary system, myocardial damage or brain damage due to hypotension may occur. Heart failure due to ischemia and myocardial injury and brain injury, for example, by bypass graft surgery during coronary artery occlusion, and aneurysm surgery in the cerebral artery or aorta. Side effects such as involuntary return may occur. Indeed, surgical or interventional treatment of aortic aneurysms causes side effects such as myocardial ischemia, kidney failure, and lower extremity palsy in 3-16% of patients. Therefore, if a drug that inhibits apoptosis in ischemic conditions is used for pretreatment, the side effects can be reduced.

The cause of neuronal suicide is not yet clear, but when the cerebral ischemia is transiently induced, the supply of oxygen and glucose is blocked, resulting in a decrease in ATP and edema in neurons, leading to neuronal suicide. Known. Neuronal suicide mechanism by cerebral ischemia is an excitatory neuronal suicide mechanism in which excess glutamate accumulates outside the cell by ischemia, and this glutamic acid enters the cell, resulting in neuronal cell death due to excessive accumulation of intracellular calcium. Kang TC. Et al., J. Neurocytol . , 30 , pp945-955, 2001) and the increase of radicals in vivo due to sudden oxygen supply during ischemia-reperfusion damages DNA and cytoplasm, causing neuronal death. Oxidative neuronal suicide mechanism (Won MH. Et al., Brain Res. , 836, pp70-78, 1999). Based on these mechanisms, many researches have been conducted to search for substances that effectively inhibit neuronal cell death caused by cerebral ischemia or to reveal the mechanism of the substance. However, few substances have been found to effectively inhibit neuronal cell suicide.

However, in the case of minocycline, a tetracycline antibiotic that inhibits apoptosis under ischemic conditions, myocardial infarction (Yrjanheikki J. et al., Proc. Natl. Acad. Sci. USA, 96 (23) , pp13496-13500, 1999 .) And ischemic diseases such as cerebral infarction (Scarabelli TM et al., J. Am. Coll. Cardiol. , 43 (5) , pp865-874, 2004.) as well as Parkinson's disease (Wu DC et al., J. Neurosci ., 22 (5), pp1763-1771 , 2002), Alzheimer's disease, peak (Pick) disease, Creutzfeldt -... Jakob (Creutzfeldt-Jakob) disease and Huntington's disease (Chen M. et al, Nat Med, 6 (7) , pp797-801, 2000), and is known to be effective in the treatment of degenerative brain diseases caused by neuronal cell suicide. In addition, the present inventors have confirmed that such tetracycline-based antibiotics improve cell survival under ischemic conditions similar to this study (US Pat. No. 6716822). Therefore, natural products that improve cell survival by inhibiting apoptosis in ischemic conditions can be inferred to be effective in treating not only ischemic diseases but also degenerative brain diseases caused by neuronal suicide.

Cheongpi ( Aurantii Immatri Pericarpium ) is a fruit of the unripe fruit of Citrus , which is known to have antitussive action, expectorant action, strengthening of the digestive tract, antibacterial action and anti-allergic effect. In addition, according to the medicinal herb made by Hyeam Hwang Do-yeon, Igigeopungsan (利 氣 祛風 散), which consists of 15 herbs including Cheongpi, was used for the treatment of stroke and guanwasa. The constituents of skin peel include flavonoids such as hesperidin, apigenin and naringin, and essential oils such as limonene.

Patents using this plant include extracts and components as cholesterol lowering agents (Korea Patent Registration No. 213895, Korea Patent Registration No. 254906, Korea Patent Registration No. 291145, and Korea Patent Registration No. 291144) ), Patents using components as a composition for inhibiting platelet aggregation (Korea Patent Registration No. 284130, Korea Patent Registration No. 298124, Korea Patent Registration No. 284129, Korea Patent Registration No. 276979), etc. In the literature, there are papers on the effect of improving the blood pressure of cheongpi (Huang YT. Et al., Life Sciences , 57 (22), pp2011-2020, 1995), but the ischemic and degenerative brain diseases of this extract There is no teaching or description in any of the above documents with regard to their use.

Accordingly, the present inventors have recognized the problems of the prior art in the treatment of ischemic disease and degenerative brain disease, and the research and development for the development of apoptosis inhibitors for improving cell viability in hypoxic conditions as a therapeutic agent for ischemic disease and degenerative brain disease. The present invention was completed by confirming the effect of the extract.

It is an object of the present invention to provide a skin extract and composition containing the same effectively inhibiting apoptosis in ischemic conditions.

In order to carry out the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of ischemic diseases containing the extract as an active ingredient.

The ischemic diseases include myocardial infarction, cerebral infarction, ischemic impaired nephropathy, diabetic foot ulcers and diseases caused by surgical side effects (heart failure, incomparable nephropathy, etc.).

The present invention provides a pharmaceutical composition for the prevention and treatment of degenerative brain diseases, containing the extract as an active ingredient.

The degenerative brain diseases include Parkinson's disease, Huntington's disease, Alzheimer's disease, Pick's disease, and Creutzfeldt-Jakob's disease.

The extract is water or an organic solvent such as a polar solvent of lower alcohols such as methanol and ethanol, a non-polar solvent of hexane, ethyl acetate, dichloromethane or a mixed solvent thereof, and preferably includes an extract available in ethanol.

Hereinafter, the present invention will be described in detail.

The skin extract of the present invention, after drying, has a volume of about 1 to 15 times the volume of the fine skin root, and preferably about 5 to 10 times the amount of C ₁ to C ₄ such as water, methanol, ethanol, and the like. Lower alcohol or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10, preferably water and ethanol, and about 0.5 hours to 2 days at an extraction temperature of 20 to 100 캜, preferably 50 to 100 캜, preferably For example, cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, herbal bath extraction, and the like are extracted for 1 hour to 1 day, preferably, 1 to 12 times, preferably 3 to 4 times, using a herbal bath extraction method. Extracted by filtration, and the filtered extract was concentrated under reduced pressure at 20 to 100 ° C., preferably at 40 to 70 ° C., using a vacuum rotary concentrator, and then the extracted residue was dried with a vacuum freeze dryer, and then water, lower alcohol, or a mixed solvent thereof. You can obtain the extracts available in.

The present invention provides a pharmaceutical composition effective for the prevention and treatment of ischemic diseases and degenerative brain diseases containing the extract of the present invention obtained as the active ingredient as an active ingredient.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical compositions comprising extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. 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, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate and 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 may include various excipients, such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to water and liquid paraffin, which are commonly used to include suspensions, solutions, emulsions, and syrups. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The compositions of the present invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. Administration may be administered once a day or may be divided several times.

The composition comprising the extract of the present invention provides a dietary supplement for the prevention and treatment of ischemic diseases and degenerative brain diseases. Examples of the food to which the extract can be added include various foods, beverages, gums, teas, vitamin complexes, and health supplements.

In addition, the extract of the present invention may be added to food or beverage for the purpose of preventing the ischemic disease and degenerative brain disease. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml. have.

Health functional food of the present invention includes the form of tablets, capsules, pills, liquids and the like.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for having the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. 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 proportion of said natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition 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 adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention will be described in more detail based on the following examples and experimental examples, but the present invention is not limited thereto.

Example 1.Preparation of Peel Ethanol Extract

After rinsing the commercially purchased skinned and washed, dried 5ml of ethanol into 500g of the skinned sample, repeated extracting twice for 24 hours in an ultrasonic extractor (sonicator), filtered, and reduced pressure with a vacuum condenser (NP-1, EYERA) Concentration gave 65 g. Filtration was carried out under reduced pressure with a vacuum concentrator (NP-1, EYRA) to obtain 50 g of skin ethanol extract.

Example 2. Preparation of Peeled Water Extract

After washing commercially, cheongpi purchased as a sample, washed with water, and 200 liters of water was added to 200g of cheongpi according to the conventional herbal extractor method, and extracted for 2 hours in an electric herbal bath (Daewoong brewing machine DWP-2000, Daewoong) and filtered. . About 2 liters of the extract obtained by filtration was lyophilized to obtain 30 g of skin extract.

Reference Example 1. Preparation of Laboratory Animals

The experimental animals were Sprague-Dawley male rats (Hyochang Science Co., Ltd.) weighing 170-230 g, and were subjected to constant conditions (temperature: 21 ± 2, contrast: 12 hours in Daegu Catholic Medical University). In the light and dark cycle), it is possible to freely consume the feed and the negative water, and provide sufficient water and food until the start of the experiment, and pre-treat the experimental animals for 10 minutes before the start of the experiment.

Experimental Example 1. Measurement of cell viability improvement capacity (in vitro)

In order to investigate the effect of the extract on the survival of cells under hypoxic and normal oxygen conditions, after adding a medium containing extracts of different concentrations to the cells, the cell survival by performing the experiment as follows under hypoxic and normal oxygen conditions The degree of improvement in the trypan blue dye exclusion described in Sambrook J. et al., Molecular Cloning 3rd ed. , 3 , pp6-8, Cold Spring Harbor Laboratory Press, 2001 It was measured by.

HepG2 (2 × 10 ⁵ cells / 800 μl), a human hepatoma cell line, was incubated in a 12 well-plate using Eagle Minimal Media. The cells were incubated for 48 hours at 37 ° C. and 5% CO ₂ -95% atmosphere, and then replaced with fresh culture medium. Then, the ethanol extract of Example 1 was dissolved in 50% ethanol and 100 µg / ml, 1000 µg. The experimental group added to the culture medium at the concentration of / ml and the negative control group (negative control) without any addition was incubated for 2 days at low oxygen (oxygen concentration 3%) and normal oxygen conditions, using the trypan blue staining exclusion method The cell number was measured.

As shown in Figure 1a, the extract improves cell survival at a concentration of 100-1000 µg / ml under hypoxic conditions and as shown in Figure 1b, the growth of cells at a concentration of 1000 µg / ml under normal oxygen conditions It was confirmed to inhibit. Accordingly, it was confirmed that the extract of the present invention effectively improves the survival of cells under hypoxic conditions and is administered at a concentration of 100-1000 ㎍ / ml to show the effect under hypoxic conditions without inhibiting cell growth under normal conditions. It could be confirmed.

Experimental Example 2. Test of the effect of inhibiting apoptosis under hypoxic condition

In order to confirm the mechanism by which the extract helps cell survival under hypoxic conditions, the DNA fragmentation assay (DNA fragmentation assay-GP Studzinski, Apoptosis , pp47-48, 1999) described in the literature was modified.

HepG2 (1 × 10 ⁶ cells / 4 ml), a liver cancer cell line, was incubated in a 60 mm dish using Eagle Minimal Media. The cells were incubated for 48 hours at 37 ° C. and 5% CO ₂ -95% atmosphere, and then replaced with fresh culture medium. Then, the extract of Example 1 was dissolved in 50% ethanol and incubated at a concentration of 400 μg / ml. The DNA fragmentation assay was performed by incubating the experimental group added to the medium and the negative control group to which nothing was added for 2 days under hypoxic conditions and normal oxygen, respectively. Investigate. As a result, the experimental group to which the extract was added was delayed while improving the survival of cells as in Experimental Example 1 as compared to the control group to which the extract was not added (see FIGS. 2A, 2B and 2C). Therefore, it was confirmed that the extract of the present invention has an effect of suppressing apoptosis and improving cell survival.

Experimental Example 3. Effect test on cerebral infarction during intraperitoneal injection (in vivo)

In order to confirm the effectiveness of the extract in the treatment of cerebral infarction (Han HS et al., Journal of Neuroscience , 22 , pp3921-3928, 2002) by modifying the experiment described in the rat of the reference example Animal experiments were performed as follows.

After inhalation anesthesia of the rat of Reference Example 1 with enflurane, the blood pressure measurement and blood sampling route were secured in the lower limb artery, and the cervical incision was exposed to expose the carotid artery. A cerebral infarction model was created by inserting 3-0 nylon sutures to block the middle cerebral artery.

The extract of Example 1 was injected into the cerebral infarction model to investigate the area of necrotic sites. At this time, the dose should be solved at the same time that it is a measurement of the safe nontoxic dose and the determination of the effective dose. Therefore, based on the result of the in vitro experiment, the method is to raise the step by step to the minimum possible starting point. Was used. To this end, the dose increase was adopted by the fold increase method, and starting at 50 mg / kg increased to 100 and 200 mg / kg.

20 hours and 1 hour before blocking the middle cerebral artery, 1 ml of 200 mg / kg of scalp were injected into the abdominal cavity, respectively, and left for 2 hours to remove the suture and the animals were recovered. After 22 hours, the animals were euthanized, brain tissues were extracted, stained in TTC solution, and the degree of damage was measured by an image analysis system to calculate the ratio of damage to the brain hemispheres. Ischemic index was measured to compare the effects of drugs.

Ischemia index (%) = A / B × 100

A: the volume of the injury site in the hemisphere of the brain (mm ³ ),

B: total volume of brain hemispheres (mm ³ ).

As a result of the experiment, rats that were not injected with dermal injection showed an ischemic index of 91% compared to rats injected with 73%, and the volume of necrotic area was reduced by 20% (p <0.05). It was confirmed that there is a significant effect (see Fig. 3a, 3b, 3c).

Experimental Example 4. Toxicity Test

The ethanol extract of Example 1 was injected into 300 Sprague-Dawley male rats of 300 ± 20 g to observe changes.

The body weight was measured 24 hours after injecting 500 mg / kg of extract of pericarp into the abdominal cavity and there was no significant weight change. There was also no change in behavioral behavior and no revenge when incised.

It describes an example of the formulation of the pharmaceutical composition containing the extract of the present invention, but the present invention is not intended to limit it, only intended to explain in detail.

Formulation Example 1 Preparation of Powder

300 mg of ethanol extract of Example 1

Lactose 100 mg

Talc 10 mg

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

Formulation Example 2 Preparation of Tablet

50 mg of ethanol extract of Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Formulation Example 3 Preparation of Capsule

50 mg of ethanol extract of Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

50 mg of skin extract of Example 2

Appropriate sterile distilled water for injection

pH adjuster

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

100 mg skin extract of Example 2

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added 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 6 Preparation of Healthy Food

1000 mg of skin extract of Example 2

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

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

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 ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

1000 mg of skin extract of Example 2

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 according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

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

Composition containing the extract of the present invention is not only to reduce apoptosis of the tissue by inhibiting apoptosis in the ischemic state, there is no side effect even in the long-term use in medicine and health functional food for the treatment of ischemic disease and degenerative brain disease Can be used.

Claims (7)

A pharmaceutical composition for the prevention and treatment of myocardial infarction and cerebral infarction, containing the extract as an active ingredient. delete A pharmaceutical composition for the prevention and treatment of degenerative brain diseases, which contains the extract as an active ingredient. The pharmaceutical composition according to claim 3, wherein the degenerative brain disease is Parkinson's disease, Huntington's disease, Alzheimer's disease, Peak disease and Creutzfeldt-Jakob disease. A dietary supplement for the prevention and improvement of myocardial infarction and cerebral infarction, which contains green skin extract and food acceptable additives. The health functional food for preventing and improving myocardial infarction and cerebral infarction according to claim 5, which is a tablet, a pill, a capsule or a liquid. delete

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