KR20050076104A - Composition for improvement of cardiovascular disease - Google Patents

Abstract

The present invention relates to a cardiovascular disease improving composition, and more specifically, 2 to 70% by weight of alginic acid derived from seaweed, 0.5 to 30% by weight of fucoidan, 1 to 50% by weight of laminarin, and 2 to phloroglucinol. It relates to a cardiovascular disease improvement composition comprising 70% by weight and 0.01 to 10% by weight of fucosterol. The composition according to the present invention has three kinds of physiological activities of hypolipidemic activity, LDL antioxidant activity, and antithrombotic activity, which can effectively block the vicious cycle of hyperlipidemia, vascular hardening and thrombogenesis, which are the main causes of cardiovascular deterioration. At the same time, there is no side effect, and has an excellent effect on improving cardiovascular disease compared to a substance depending on one activity.

Description

Composition for improvement of cardiovascular disease

The present invention relates to a cardiovascular disease improving composition, and more particularly, cardiovascular system using a bioactive material comprising polysaccharides, fluoroglucinols and fucosterols composed of alginic acid, fucoidan and laminarin derived from seaweeds. In the cardiovascular disease improving composition that effectively suppresses and improves the vascular hardening factor caused by the increase of blood cholesterol, which causes the disease, the vascular wall destruction caused by the low-density lipoprotein (LDL) oxidation, and the deterioration of blood flow due to the formation of blood clots. It is about.

The first condition to improve cardiovascular disease is to normalize excessively elevated blood LDL cholesterol levels to block cardiovascular deterioration factors, and the second condition can block vascular hardening even under elevated blood cholesterol conditions. The third condition is to promote the inhibition and degradation of thrombus formation, which is likely to be produced in the exacerbated vasculature. If only one of the three conditions is met, it may be effective in the prevention and improvement of cardiovascular diseases, but if the use of drugs with high toxicity or side effects, or if the cardiovascular disease has already advanced considerably, the practical effect may be insignificant. have. That is, when the three conditions such as thrombolysis, LDL oxidation prevention, and cholesterol removal are satisfied at the same time, the cardiovascular disease improvement effect may be maximized.

The coagulation and dissolution of the blood is always in equilibrium in vivo, so in normal conditions, blood flow is not disturbed by bleeding or blood clots. However, when this equilibrium is broken by various factors, blood flow is not smooth, which causes cardiovascular disease. In general, the formation of clots in the active cardiovascular system is called thrombosis (Thrombosis), the clot is called a blood clot (Thrombus). These clots interfere with the circulation of blood vessels in veins and arteries, blocking the supply of nutrients and oxygen to tissues, leading to serious adult diseases such as cerebral hemorrhage, heart failure myocardial infarction, and atherosclerosis.

For these reasons, many studies have been conducted to treat thrombi. Specifically, plasminogen activators such as streptokinase and urokinase have been known to be useful for removing blood clots, and therapies for activating the fibrinolytic system by intravenous injection have been known for the past 30 years. Has been used. The thrombolytic effect of the two drugs has already been demonstrated in clinical trials, but side effects such as systemic bleeding occur during treatment. Recently, attempts to overcome the problems of the thrombolytic agent have been very active. For example, the development of new mutant activators with strong thrombolytic activity and development of new thrombolytic agents derived from animals, plants or microorganisms with better thrombolytic ability from natural substances are in progress.

In particular, studies using plants are very active, and many studies on tissue factor pathway inhibitors (TFPI) have been reported. Specifically, ingredients extracted from ginkgo biloba are known to facilitate blood circulation, and formulations using the same have been developed and commercialized. A blood clot soluble substance has been reported in animals. For example, a new plasminogen activator (Gardell, SJ et al., 1989) isolated from the bloodsucking bat tissue of the Merck Institute in the United States. , (1991) Circulation, 84, 244-253), salmonase isolated from snake venom (Chung, KH and Kim, DS, (1991) Thromb. Haemost., 65, 953), isolated from leeches Hementin and fibrinolytic enzyme isolated from centipedes (Kim, KY et al., (1993) Thromb. Haemost., 69, 83) have been reported as novel thrombolytic agents. In addition, research on thrombolytic enzymes in fermented foods that can be directly consumed is being conducted in Korea and Japan.

Aerobic organisms obtain energy through respiration using oxygen as an electron acceptor. As such, oxygen is absolutely necessary for maintaining life, but when oxygen is converted into active oxygen in the body due to various stresses or food additives or environmental pollution, it causes fatal oxygen toxicity to the living body. That is, lipid peroxides produced by peroxidation of lipids by these free radicals and various peroxides in the body oxidatively destroy cells, causing various dysfunctions, which causes aging and degenerative diseases. In normal cells, some free radicals and other free radicals and peroxides are produced during metabolism, but in general, superoxide dismutase (SOD) and catalysts are used as a defense against them. Along with antioxidant enzymes such as catalase and peroxidase, antioxidants such as vitamin E, vitamin C, glutathione, ubiquinone, and uric acid exist to protect themselves. However, when an abnormality occurs in such a biological defense mechanism or the generation of free radicals exceeds the capacity of the biological defense system by various physical and chemical factors, oxidative stress occurs. Therefore, antioxidants such as free radical scavengers or peroxide production inhibitors are essential to suppress cardiovascular diseases caused by the oxides. Antioxidants which have been developed and used so far include synthetic antioxidants such as tert-butylhydroxytoluene (BHT) and tert-butylhydroxyanisole (BHA), and alpha-tocopherol. , Some natural antioxidants such as vitamin C, carotenoids, flavonoids and the like and antioxidant enzymes such as SOD are limited. However, these antioxidants are limited to use due to various problems such as toxicity, low activity and limit of use. Since the late 1980s, there have been attempts to extract physiologically active substances, especially antioxidants, from seaweeds, and much research has been conducted especially in France and Japan. Takaki and Miyashida extracted natural products from 12 kinds of seaweeds in the waters of Japan, and investigated the tocopherol component of these substances. (K. Miyashita and T. Tagaki, Agric. Biol. Chem. 51, 315 (1987)). Nishibori and Nakami extracted antioxidants from hexane / ethanol mixtures of 7 kinds of seaweeds, and measured their antioxidant activity. And antioxidant activity comparable to alpha-tocopherol (S. Nishibori and K. Namiki, Home Economics Magazine 36, 17 (1985)). However, this too was difficult to commercialize due to the very low extraction amount. In Korea, Park Jae-han et al. Obtained 12 extracts of algae using methanol and chloroform to measure their antioxidant activity. As a result, we have obtained better antioxidants than BHA in seaweed, seaweed, and kelp, but have not been commercialized due to thermal stability (Jae-han Park, Kyu-chan Kang, Sang-bong Baek, Yun-hyeong Lee, Kyu-soon Lee, Korean Food Science Society 23, 256 ( 1991).

As such, oxidative stress such as lipid peroxidation indiscriminately destroys tissues and blood cells, and has been proven to be an important cause of hyperlipidemia and cardiovascular disease by degrading blood flow and blood quality through the formation of blood clots and hardening of blood vessels. The possibility of LDL antioxidant compositions using an antioxidant component having free radical scavenging activity has been greatly highlighted. However, natural and synthetic antioxidants have been developed and used as medicines and various health supplements to date, but there are limitations in preventing cardiovascular diseases due to oxidation of LDL due to various problems in toxicity and in vivo.

Prolonged hyperlipidemia causes arteriosclerosis in blood vessels through which blood is circulated. When such an arteriosclerosis occurs, blood circulation to the brain or heart may not be a life threat. Atherosclerosis is caused by excessive deposition of low-density lipoprotein-cholesterol that is increased in the blood inside the arteries, and inflammatory cells such as leukocytes are gathered here and smooth muscle cells proliferate. The deposition of lipids inside the arteries occurs with hyperlipidemia, even when the vessel wall is damaged by hypertension and when high-density lipoprotein-cholesterol is reduced. Atherosclerosis develops, blood vessels narrow gradually, elasticity decreases, and blood flow decreases. A decrease in blood flow lowers the supply of oxygen and nutrients to many organs, and beyond certain thresholds, ischemic cardiovascular disease or stroke can occur. Atherosclerosis is caused by hyperlipidemia, but when hypertension, diabetes mellitus, and smoking habits are combined, the blood vessel walls are further damaged, thereby further accelerating the development and progression of atherosclerosis, and increasing the risk of cardiovascular disease. Therefore, ischemic heart disease occurs a lot in people with high cholesterol in the blood, and high cholesterol, in the case of people who smoke or have high blood pressure, ischemic cardiovascular disease occurs more. In clinical practice, vascular stenosis is suppressed by prescribing long-term use of statin cholesterol lowering agents, anticoagulants such as COUMADIN, and antiplatelets such as aspirin. 3: 1-12,1996). Thrombolytic agents, such as urokinase and tissue-type plasminogen activator (t-PA), are used as effective therapeutics to help the flow of blood by dissolving already formed clots. However, as an antiplatelet agent, aspirin is effective, but side effects such as gastrointestinal disorders have been reported, and anticoagulant coumadine has a side effect of bleeding due to its strong activity, and thus it is limited to use as a long-term primary preventive agent. There is. In addition, the statin-based drugs used for the treatment of hyperlipidemia have shown a very good effect to lower the low-density lipoprotein in the blood, but there is a problem that the price is too expensive. After all, the administration of these effective drugs is a second-order prescription for patients who have already developed cardiovascular disease, and an effective primary preventive agent for the underlying prevention has not been developed yet.

Therefore, there is a need for the development of a composition for the improvement of cardiovascular diseases that can solve the problems with existing therapeutic agents without side effects.

Accordingly, the present inventors conducted a study to solve the problems with the conventional cardiovascular disease improvers, as a result, alginic acid derived from alginic acid, fucoidan and laminarine polysaccharides, phloroglucinol and fucosterol including fucosterol Cardiovascular disease improvement composition that effectively suppresses and improves the vascular hardening factor caused by the increase of blood cholesterol, which causes the cardiovascular disease, the vascular wall destruction by LDL oxidation, and the deterioration of blood flow due to thrombus formation. Developed. As a result of clinical trials, the composition acts simultaneously on hypolipidemic activity, LDL antioxidant activity and antithrombotic activity, which can effectively block the vicious cycle of hyperlipidemia, vascular hardening and thrombogenesis, which are the main causes of cardiovascular deterioration. Compared with a substance without any activity, it showed an excellent effect on improving cardiovascular disease, and it was economically, safe, and convenient to consume in the form of food as well as medicine, and the present invention was completed based on this. It became.

Accordingly, an object of the present invention is to lower blood cholesterol by using a bioactive substance including polysaccharide, algal acid made of algae, fucoidan and laminarin, phloroglucinol, and fucosterol, thereby causing hyperlipidemia and abnormal blood function. It is possible to prevent the onset of various cardiovascular diseases, and has a strong antioxidant component that can inhibit free radicals, which is the main cause of destruction of vascular endothelial cells due to peroxidation of LDL and the formation of blood clots. Cardiovascular disease improving composition that can effectively prevent and improve vascular circulatory diseases such as arteriosclerosis, hypertension, ischemic heart disease, and stroke by having components that can inhibit thrombolysis, platelet aggregation activity and excessive coagulation tendency To provide.

The composition of the present invention for achieving the above object is 2 to 70% by weight of alginic acid derived from seaweed, 0.5 to 30% by weight of fucoidan, 1 to 50% by weight of laminarin, 2 to 70% by weight of phloroglucinol and fucosterol 0.01 to 10% by weight.

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

As described above, the present invention relates to a cardiovascular disease improving composition, and more particularly, to a bioactive material including polysaccharides consisting of alginic acid, fucoidan, and laminarin derived from seaweed, fluoroglucinol, and fucosterol. Cardiovascular system effectively inhibits and improves blood vessel hardening factors caused by an increase in blood cholesterol, which causes cardiovascular disease, vascular wall destruction caused by low-density lipoprotein (LDL) oxidation, and blood flow deterioration caused by blood clots. A relationship disease improving composition.

In general, many seaweeds used for food use are bioactive substances, and polysaccharides such as fucoidan, anti-blood coagulation, laminarin, blood pressure lowering, alginate, cholesterol lowering, and waste elimination, and various forms of bioactivity. It contains the ingredients: phloroglucinol, which has potent antioxidant and thrombolytic functions, and fucosterol, which has a cholesterol-lowering effect. Therefore, the composition of the present invention is characterized by the optimal combination of seaweed-derived bioactive substances.

The alginic acid used in the present invention is a polymer of two kinds of uronic acids (mannuuronic acid and gluuronic acid), and the brown algae are washed with dilute sulfuric acid, extracted from dilute alkaline hot water, and then acidified. This is alginic acid. Alginic acid is mostly present in combination with potassium, sodium and calcium. When potassium alginate enters the stomach, potassium falls by the action of stomach acid. Since the intestines are weakly alkaline, alginic acid easily binds to minerals when they enter the intestines. Alginate binds much with sodium because there is a lot of sodium in the intestine. Alginic acid is difficult to digest and is not absorbed in the intestine, so potassium alginate eventually becomes sodium alginate and is excreted with feces. Potassium released from alginic acid, on the other hand, is absorbed by the intestine and replaced with sodium in the blood. Potassium lowers blood pressure because it has an action to lower blood pressure. In addition, alginate is a pectin-like substance, and has a strong gel-forming property, which is one of the factors of cholesterol lowering effect, so that the function of lowering blood cholesterol is known (Ray H. Crist and J. Robert Martin, Environmental Science). and Technology, 1999, 33, 2252-2256).

The amount of the alginic acid is 2 to 70% by weight, preferably 20 to 50% by weight, based on solid content, less than 2% by weight is difficult to see the effect of cholesterol removal, more than 70% by weight of anti-thrombolytic or LDL oxidation prevention There is a tendency to reduce the effectiveness of the composition for playing a role. In addition, the molecular weight of the alginic acid is 3,000 or more, the average molecular weight is 15,000 to 150,000, preferably the molecular weight is 20,000 or more, the average molecular weight is 30,000 to 120,000 is preferred in order to obtain the absorption and optimum efficacy.

In addition, the fucoidan (Fucoidan) contains galactose, xylose, glucuronic acid (Glucuronic acid) and the like as a main component of esterified sulfuric acid of L-Fucose (L-Fucose) As polysaccharides, sulfated fucose-containing polysaccharides are collectively referred to. Fucoidan (sulfated fucose-containing polysaccharides) is a class of heparinoids known to have anticoagulant effects (Carbohydrate Research, 1991, 211, 77-90, Carbohydrate Research, 1991, 214, 193-197). , Carbohydrate Research, 1992, 229, 355-362).

The amount of fucoidan is 0.5 to 30% by weight, preferably 2 to 20% by weight, based on solid content, less than 0.5% by weight is difficult to expect the effect of preventing blood coagulation, when exceeding 20% by weight exhibits optimal synergy There is a tendency to make the composition to do it. The molecular weight of the fucoidan is 5,000 or more, the average molecular weight is 20,000 to 200,000, the sulfated content is 5 to 45%, preferably the molecular weight is 15,000 or more, the average molecular weight is 20,000 to 150,000, the sulfated content is 10 to 25%. Required for absorption and metabolism in the body.

On the other hand, the laminarin is a storage polysaccharide equivalent to the starch of higher plants, and there are soluble laminarin and insoluble laminarin, which are composed of β-1,3-glucan, but the former is a small number of β-1,6- It also forms glycosidic bonds and is known to have a good effect on blood pressure lowering.

The amount of laminarin is 1 to 50% by weight, preferably 5 to 40% by weight, based on solids, less than 1% by weight of the blood pressure lowering function is insignificant, more than 50% by weight to improve the cardiovascular system Formulation for optimization is difficult

The phloroglucinol is contained in brown algae marine plants and has excellent antioxidant activity (J. of Korean ind. & Eng. Chemistry, 1996, 7, 1069-1077). The phloroglucinols are superior to synthetic antioxidants such as BHT and BHA, have superior antioxidant activity as compared to natural vitamin E or C, and are very resistant to heat and are hardly destroyed in processing. One of these components, dieckol, has a dibenzo-p-dioxin structure and is known to activate the thrombolytic action of plasmin by inhibiting the action of anti-plasmin (Anti-Plasmin) ( Chem. Pharm. Bull. 1989, 37, 349-353).

The amount of phloroglucinol used is 2 to 70% by weight, preferably 10 to 50% by weight, and less than 2% by weight, it is difficult to express the effects of thrombolysis and LDL oxidation, and 70% by weight. If exceeded, the formulation between the compositions that can maximize the improvement of cardiovascular disease can not be made properly. The phloroglucinols include ecol, dieckol, phlorotannin A, phlolofucopuroecol, 7,7'-bieckol, 9,9'-bieckol and glycosides thereof.

Fucosterol, on the other hand, is called plant sterol or plant stanol, which is structurally similar to cholesterol synthesized in animals, and in the cell membranes of plants, just as cholesterol is an essential ingredient in plant cell membranes. This is one of the essential ingredients. Vegetable stanols are in the form of saturated sterols. Representative phytosterols include fucosterol, beta-sitosterol, beta-sitostanol, campesterol, stigmasterol and the like. Clinically, sterols and stanols are used to treat cholesterol-lowering effects in patients with hyperlipidemia. Serum total cholesterol levels range from 5 to 23 as a result of feeding phytosterol blends or phytosterol-sourced foods to patients with hyperlipidemia. %, Serum LDL cholesterol was reduced by about 8-29% (The American Journal of Medicine, 2000, 109, 600-601).

The amount of fucosterol is 0.01 to 10% by weight, preferably 0.05 to 5% by weight, based on solids, less than 0.01% by weight is difficult to expect the effect of lowering cholesterol, more than 5% by weight improve cardiovascular disease The formulation between the compositions to maximize the can not be made properly.

The bioactive substance is brown seaweed, mabanban, forked bear, green, Ecklonia, sesame, kelp, rhubarb, laver, sea stalk, stone starfish, round stone, steamed, falconus, independence starfish, stone, spirulina, wood starfish, jindubal, hearing, One or more selected from the group consisting of methanol, ethanol, ethyl acetate, acetonitrile, ether, acetone, chloroform, hexane, water and mixtures thereof and water / ethanol mixtures from one or more selected seaweeds selected from the group consisting of chlorella and gompi Obtained by the method of extracting and refine | purifying using the solvent chosen above.

 In addition, the composition may be added to medicines, dietary supplements, special nutritional products, beverages and alcohol, food additives, sweets or bread alone or in the form of the combination.

The daily dose of the composition of the present invention is 2g or more per 1kg body weight, and even if ingested in excess of the product obtained from natural foods, there are no side effects.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the scope of the present invention is not limited to the following Examples.

The effective bioactive substances contained in the seaweeds are different and the extraction methods are varied, and it is difficult to prepare effective composition for improving cardiovascular disease with a single seaweed, so the bioactive substance as the main ingredient is extracted as described below in an optimal economic manner. It was. The selection of algae was based on the anticoagulant, antithrombotic activity and antioxidant power, and various folk remedies and clinical data of algae under laboratory conditions, but the separation of certain bioactive substances is limited to the specific algae shown in the examples below. It is not. That is, the bioactive substance for the cardiovascular disease improving composition can be obtained by extracting all terrestrial and marine natural plants as raw materials.

Example 1

Isolation of Alginic Acid from Kelp

Kelp was dried with tap water and dried in a well-ventilated place. 300 g of dried kelp was finely crushed using a blender, put into 3000 ml of distilled water, stirred under reflux for 1 hour, and separated into an aqueous solution and an insoluble precipitate. After dissolving the undissolved precipitate in 3000ml of distilled water and refluxing for 1 hour, the mixture was separated into an aqueous solution and an insoluble precipitate, and the aqueous solution was combined with the first aqueous solution and lyophilized to obtain 125 g of alginic acid powder.

Example 2

Isolation of Fucoidan from Gompi

Gompi was dehydrated with tap water and shaded in a well-ventilated place. 300 g of dried Gompi was finely crushed using a blender and placed in 3000 ml of distilled water and stirred under reflux for 1 hour, and then separated into an aqueous solution and an insoluble precipitate. The undissolved precipitate was added again to 3000 ml of distilled water and stirred under reflux for 1 hour, and then separated into an aqueous solution and an undissolved precipitate. The aqueous solution was combined with the first aqueous solution and lyophilized to obtain 75 g of powder. 75 g of the powder was dissolved in 30 ml of distilled water, and then precipitated by addition of an aqueous 5% CaCl ₂ solution. The solution was centrifuged to concentrate the supernatant under reduced pressure, and high molecular weight fucoidan was separated using a separator. The component in the separator was fucoidan, which was lyophilized to obtain 1.2 g.

Example 3

Isolation of Laminarin from Seaweed

Seaweed was dried in a well-ventilated area by tapping with salt water. 300 g of dried seaweed was finely crushed using a blender, placed in 1000 ml of distilled water, stirred at room temperature for 2 hours, and an aqueous solution and an insoluble precipitate were separated. The separated precipitate was added to 2000 ml of distilled water, and the mixture was stirred under reflux for 2 hours. The precipitate was separated into an aqueous solution and an undissolved precipitate, and the aqueous solution was lyophilized to obtain 5.6 g of laminarin powder.

Example 4

Isolation of Phloroglucinol from Ecklonia cava

Ecklonia cava was shaded in tap water to remove salt. 300 g of dried Ecklonia cava was finely crushed using a blender and placed in 3000 ml of 95% alcohol, stirred at room temperature for 24 hours, and then separated into an alcohol layer and an insoluble precipitate. The alcohol layer was dried under reduced pressure to obtain 30 g of powder. 30 g of the powder was added to 300 ml of absolute ethanol, and stirred for 1 hour. The undissolved precipitate was separated, and the ethanol layer was dried under reduced pressure to obtain 18 g of a compound having a concentrated phloroglucinol component.

Example 5

Isolation of Fucosterol from the Mother Board

After removing salt with tap water and lyophilizing, 300 g of dried mabanban is finely crushed with a blender and placed in 3000 ml of chloroform / methanol (2/1) solution and stirred at room temperature for 24 hours. Separated into precipitate. The organic layer was dried under reduced pressure to obtain 35 g of powder. 35 g of this powder was hydrolyzed with 10% KOH for 4 hours using 600 ml of ethanol as a solvent. After the reaction solution was cooled, 1000 ml of water was added thereto, followed by extraction with ether to obtain 5.1 g of a compound having a concentration of fucosterol.

Example 6

Preparation of Seaweed Complex Group for Determination of Cardiovascular Disease Improvement Composition Using Seaweed Extract Bioactive Materials

A single seaweed or a single bioactive substance cannot play a role in LDL oxidation inhibition, thrombolysis, and cholesterol removal, which are essential for the suppression and improvement of these cardiovascular diseases. Therefore, antioxidant activity to prevent LDL oxidation, reduction of blood cholesterol and waste products, and thrombus In order to prepare a cardiovascular improvement composition having an activity of dissolution, five bioactive substances extracted through the above examples were combined as shown in Table 1 below. Various combination ratios of the five bioactive substances may come out, but the following five complex groups were selected in consideration of efficacy and economic efficiency of extraction. According to the following content ratio, the five bioactive substances were uniformly mixed using a blender.

Ingredients and Contents of Seaweed Extracted Bioactive Substances Compound ingredient A B C D E Phloroglucinol 32 g 0g 32 g 32 g 32 g Fucoidan 6 g 6 g 0 6 g 6 g Alginic acid 25 g 25 g 25 g 0 25 g Laminarin 20 g 20 g 20 g 20 g 0 Fucosterol 0.1g 0.1g 0.1g 0.1g 0

Example 7

Anticoagulant and Antithrombotic Activity and Antioxidant Activity by Component Ratio

1 g each of the algae complex group of Example 6 was tested for anticoagulant and antithrombotic activity and antioxidant activity, and an experiment using a rat was performed to compare the drop of cholesterol.

In order to measure activated partial thromboplastin times (aPTT), samples were first dissolved in an appropriate amount of DMSO and diluted with distilled water so as to have a concentration of 3.3 mg / ml. 50 μl of sample and 100 μl of plasma were mixed well, and then 50 μl of the sample was measured for coagulation time (final sample concentration of about 367 ppm during analysis). The anticoagulant activity% was expressed by multiplying the coagulation time (sec) at the time of sample addition by the coagulation time of the control group by 100, wherein the control group was 54.6 seconds. Heparin was used as a positive control. The aPTT measurement results are shown in Table 2 below.

Thrombin times (TT) assay dissolves 1 vial of thrombin reagent (3-4 NIH units) in 5 ml distilled water and 1 vial of fibrinogen reference reagent (250 mg / dL) in 4 ml distilled water. After thawing, 50 μl of thrombin reagent and 50 μl of sample are incubated for exactly 2 minutes, and then 100 μl of fibrinogen reference reagent is added to measure the solidification time (final sample concentration is about 550 ppm). At this time, the coagulation time of the control group added distilled water instead of the sample was 61.2 seconds, and the anticoagulant activity was expressed in% by the same method as the aPTT measurement. The experiment was repeated three times, and the results are expressed as their average values, which are shown in Table 2 below.

Antioxidant activity was measured using the bios method in vitro method as follows. Approximately 20 mg of 1,1-diphenyl-2-picrylhydrazyl (DPPH, Aldrich) was dissolved in 150 ml of ethanol to prepare DPPH solution. Then, 250 ml of dimethyl sulfoxide (DMSO) was added, diluted with an appropriate amount of ethanol and shaken for 10 seconds. And the concentration was adjusted so that the absorbance of a control group might be 0.94-0.97 at the wavelength of 518 nm. In the same manner, 100-200 μl of the sample (1 mg / mL) solution was added to 3 mL of DPPH solution having an absorbance of 0.94-0.97, respectively, and the mixture was sufficiently mixed, followed by reaction for 10 minutes. Then, the absorbance of the reactants was measured to show the antioxidant activity as the DPPH radical scavenging activity of the absorbance decrease for the control group, which is shown in Table 2 below.

Anticoagulant and Antithrombotic Activity and Antioxidant Degree by Component Ratio aPTT TT Antioxidant levels Compound group A 392 286 0.836 Compound Group B 18 13 0.041 Compound Group C 24 38 0.764 Compound Group D 86 115 0.726 Compound group E 316 235 0.731

Example 8

Cholesterol and Lipid Lowering Comparison

In order to measure the concentration change of total cholesterol, free cholesterol and cholesterol ester for the seaweed composite group of Example 6, SD rats (Sprague-Dawley rat) to prepare a synthetic diet for hyperlipidemia It was ingested for 6 weeks and the dietary intake and body weight of the SD rats were measured once a week. The components and the content ratio of the synthetic diet are shown in Table 3 below.

 Composition and Content Ratio of Hyperlipidemic-Induced Synthetic Diet ingredient content Sugar Vitamin-Free Casein Inorganic MixturesVitamin MixturesLedCholesterolChlorine ChlorideAlphacel 65% by weight 15% by weight 3.5% by weight 1% by weight 2% by weight 8% by weight 1% by weight 0.05% by weight 5.45% by weight  Sum 100 wt%

The experiment was divided into a normal group ingested only a general feed, a control group ingested only a hyperlipidemic-induced synthetic diet, and each administration group orally administered the synthetic diet and the seaweed complex group of Example 6 by 100 mg / kg daily. In addition, the administration group which ingested said complex groups A-E corresponds to administration group A-E, respectively. Blood collections were fasted 12 hours prior to rule out changes in diet. At the time of blood collection, the blood was opened under ether anesthesia, blood was collected from the abdominal aorta into a test tube containing anticoagulant, and then, plasma was collected by centrifugation at 3000 rpm for 15 minutes, and stored frozen until biochemical analysis. The contents of total cholesterol, free cholesterol and cholesterol esters of the collected plasma were analyzed, and the results are shown in FIGS. 1, 2 and 3, respectively.

As shown in FIG. 1, the total cholesterol level was 2.51 times higher in the hyperlipidemia-induced dietary intake control group (212.3 mg / dl) than in the normal group (84.6 mg / dl), and the cholesterol ester value was 63.3 mg / dl to 186.1 mg / dl. Although the increase was about 3 times, the free cholesterol value was observed to be almost unchanged. As a result, it was confirmed that hyperlipidemia induction for the hyperlipidemia induced diet was well performed. Among the groups treated with the hyperlipidemia-induced diet and seaweed complex group, the group A (administered group A) had a total cholesterol level of 136.3 mg / dl, which was about 36% lower than that of the control group, and the cholesterol ester level was 186.1. The reduction effect of about 38% was shown in the mg / dl ~ 115 mg / dl. The measurement of free-cholesterol did not show a significant difference in both normal, control and administration groups. This shows that seaweed complex group A is most effective in suppressing cholesterol.

In the measurement of lipid changes in blood, triglycerides obtained in the plasma were subjected to diagnostic reagents according to the Buccolo method (Bucolo G., et al., Clin. Chem., 19: 456, 1973). The absorbance was measured at 550 nm, and blood total cholesterol was measured by colorimetry by measuring the absorbance at 500 nm using the enzyme method. Determination of high density lipoprotein (HDL) cholesterol was determined by precipitating and removing cholesterol except HDL-cholesterol in plasma, and measuring the supernatant in the same way as total cholesterol, and low density lipoprotein (LDL) -cholesterol was measured. Only the LDL-cholesterol in the plasma was precipitated as a separate solution and measured in the same manner as the total cholesterol. Atherosclerosis index was calculated by subtracting HDL-cholesterol from total cholesterol and subtracting HDL-cholesterol.

As described above, animal experiments were conducted to determine the changes in HDL-cholesterol and LDL-cholesterol. The results showed that LDL-cholesterol was a hyperlipidemia-induced dietary intake control (72.5 mg / dl) for the normal group (12.3 mg / dl). Although increased 5.9-fold (see Figure 4), HDL-cholesterol showed a slight decrease (see Figure 5), indicating that hyperlipidemia-induced diets rapidly increased LDL-cholesterol in the blood and did not affect HDL-cholesterol content. Is showing. After administration of the seaweed complex group for 6 weeks, the HDL-cholesterol and LDL-cholesterol concentrations were observed. HDL-cholesterol showed little change in all 5 administration groups (see FIG. 4), but LDL-cholesterol levels were administered in Group A (see FIG. 4). 47.3 mg / dl) showed a significant decrease of 35% compared to the control group (72.5 mg / dl) fed the hyperlipidemia induced diet only (see FIG. 5). As a result of measuring the HDL / LDL composition and the arteriosclerosis index, the HDL / LDL composition ratio was 0.108 in the control group and the administration group A was 0.228, which was about 2.1 times higher than the control group (see FIG. 6). As a result of calculating the atherosclerosis index based on the above results, the control group was 26.2, whereas the administration group A showed 11.0 as the administration of the seaweed extract complex group significantly lowered the atherosclerosis index (see FIG. 7). In triglyceride content, which is triglyceride, the concentration of triglyceride was decreased by about 24% from 78 mg / dl to 59 mg / dl in the administration group A than the control group (see FIG. 8). This suggests that Complex Group A significantly affects metabolism of triglycerides as well as cholesterol metabolism. In order to easily compare the above numerical values, they are shown in Table 4 below.

Numerical comparison table of hyperlipidemia diet group Analysis group Total Cholesterol (mg / dl) Free Cholesterol (mg / dl) Cholesterol ester (mg / dl) HDL Cholesterol (mg / dl) LDL cholesterol (mg / dl) HDL / LDL composition ratio Arteriosclerosis index Triglycerides (mg / dl) Normal Army Control Group A Group B Group B Group C Group D Group E   84.6212.3130.1172183161156 21.326.221.32324.621.822.1   63.3186.1129157149166132 19.2 7.810.8 9.2 9.6 8.9 9.4 12.372.547.352576455 1.5610.1080.2280.1770.1680.1390.182   3.4126.211.017.718.117.115.6 44.2785966686964

In summary, it can be said that the seaweed complex group A has an optimal combination for the improvement of cardiovascular disease. Therefore, the following example was implemented with the composition containing complex group A. FIG.

Example 9

Toxicity test of cardiovascular disease improving composition

In order to investigate the toxicity when repeated oral administration of the cardiovascular disease improvement composition using seaweed extract bioactive substances, the media control and the test substance (Compound Group A) were both 2000 mg / kg and 667 mg / kg, respectively. And 10 SD male and female rats were repeatedly orally administered at a dose of 222 mg / kg for 4 weeks. The test substance was measured and suspended in a 0.5% carboxymethylcellulse sodium (CMC-Na) solution, which was a solvent, to prepare the highest dose of test substance, and the low dose of the test substance was diluted step by step with the highest dose of test substance. It was prepared by the method. In the administration method, the animals were fixed by the cervical skin fixation method and injected directly into the stomach using a metal oral administration sonde. The prepared test substance was administered after suspension well using a tube mixer immediately before the start of administration. The test substances were administered once daily for 4 weeks, the body weight was measured immediately before the start of administration, and the body weight was measured every week to calculate the dose. The amount of administration was calculated at 10 ml / kg.

No toxicological changes were observed when the oral administration of the cardiovascular disease improving composition using the bioactive substance to the rat was repeated orally for 4 weeks. Therefore, the daily intake of the cardiovascular disease-improving composition using the seaweed extract bioactive substance was determined to be more than 2000mg / weight 1kg / day.

Example 10

Determination of LDL oxidation inhibition according to the concentration of cardiovascular disease improving composition (TBARS measurement)

100 μl of LDL (5 mg / mL) and 100 μl of the prepared sample were added together, and 800 μl of 30 μM CuSO 4 (in the presence of phosphate buffer solution (pH 7.4)) was added so that the total volume was 1 mL, followed by 37 ° C. water bath) for 4 hours. Then, 300 μl of 100 uM EDTA was added to stop the oxidation of LDL. 1 ml of TCA-TBA-HCl (reactant) was added to 500 µl of the oxidized LDL solution, heated at 90 ° C. for 30 minutes, cooled, and centrifuged to obtain a supernatant, and the absorbance was measured at 535 nm. The TCA-TBA-HCl reaction was prepared by mixing 15% v / v trichloroacetic acid, 0.375% v / v thiobarbituric acid and 0.25N hydrochloric acid.

The oxidation degree of LDL measured by the absorbance is shown in FIG. 9. As can be seen in Figure 9, when the degree of oxidation of the LDL in the state without the cardiovascular disease improving composition 100%, it can be seen that the higher the concentration of the cardiovascular disease improving composition, the lower the oxidative power of LDL. . Therefore, the cardiovascular disease improving composition of the present invention can be said to be excellent in the ability to inhibit LDL oxidation, which is an essential condition for the prevention and improvement of cardiovascular diseases.

Example 11

Determination of Blood Anticoagulant Activity of Cardiovascular Disease-Ameliorating Compositions

For blood anticoagulant activity, the cardiovascular disease-improving composition comprising the composite group A according to the present invention was compared with the extract of ginkgo biloba extract, which has been proven effective as a blood circulation improving agent. The experimental method was repeated three times using the aPTT assay described above, and the results of blood coagulation activity are expressed as their average values. The control group added distilled water instead of the sample showed 61.2 seconds, 71.2 seconds when the ginkgo biloba extract was added, and 392 seconds when the cardiovascular disease improvement composition according to the present invention was added (see FIG. 10). Through the above results, it was confirmed that the cardiovascular disease improving composition of the present invention exhibited a blood coagulation inhibitory effect more than five times than the extract of Ginkgo biloba.

Example 12

Activity measurement of anti-α ₂ -plasmin inhibitor (α-PI)

The activity of the anti-α ₂ -plasmin inhibitor (α-PI) was compared with the bieckol which is known to have good inhibitory activity of the composition of the present invention and the plasmin inhibitor.

Experimental method was prepared by adding 3 μg of α-PI and the sample material to 0.8 ml of 0.05M tris-hydrochloric acid buffer solution (containing pH 7.4, 0.1 NaCl) at each concentration and incubating for 20 minutes at 37 ℃, and then 0.1M sodium phosphate buffer solution. 0.05 unit of plasmin in 0.1 ml (pH 7.4, containing 25% glycerin (v / v)) was added. After incubating the mixture at 37 ° C. for 30 seconds, 0.1 ml of 3 mM S-2251 was added. Then, after incubation at 37 ° C. for 30 minutes, absorbance at 405 nm was measured by adding 0.1 ml of 50% acetic acid, which is shown in Table 5 below compared with Bieckol.

Inhibition rate for the α ₂ -plasmin inhibitor and IC ₅₀   Concentration (µg / ml)   6,6'-bieckol Compositions of the Invention 0.1 35 31 0.5 52 40 1.0 65 45 5.0 79 72 10.0 89 80 IC ₅₀ (μg / ml) 0.43 1.44

As can be seen in Table 5, the composition of the present invention can be seen that the anti-α ₂ -plasmin inhibitor activity more than three times higher than the bieckol is the optimal composition for thrombolysis.

Example 13

Effect on the human body when ingesting the cardiovascular disease improving composition according to the present invention

In order to determine the effect on the human body when ingesting the cardiovascular disease-improving composition according to the present invention, 11 patients with coronary atherosclerosis, hyperlipidemia (total constriction) of which at least one blood vessel was narrowed by at least one blood vessel on coronary angiography was confirmed. Cholesterol 220 mg / dl or LDL cholesterol = 130 mg / dl) Twelve middle-aged men and 18 postmenopausal hyperlipidemic women were examined for changes in serum lipid and lipoprotein levels and vascular endothelial dilatation function of the brachial artery. In the middle-aged female group, the experiment was conducted for 6 weeks, the middle-aged male and male patients group for 8 weeks, and 300 mg of the cardiovascular disease-improving composition according to the present invention was taken three times a day. The test was conducted at 6 weeks (female group) and 8 weeks (male group and male patient group) at the end of the experiment.

As a result of the above clinical trial on the cardiovascular disease improvement composition according to the present invention, the triglyceride in the blood was reduced about 9% from 215 ± 46 mg / dl to 195 ± 50 mg / dl, and total cholesterol was 240 ± 11 mg / dl. About 12% to 211 ± 9 mg / dl. LDL cholesterol was reduced by about 15% from 153 ± 10 mg / dl to 131 ± 8 mg / dl. No changes in the concentrations of apolipoproteins A1 and B were observed, but HDL cholesterol was 59 mg / dl at 59 mg / dl. About 11% improvement in mg / dl. The results of the clinical trials were calculated for 39 people by stopping 2 people during the experiment, which is shown in Table 6 and FIG.

Changes in Serum Lipid and Lipoprotein Concentrations after Ingestion of Cardiovascular Disease Improvement Composition N = 39 Before intake After ingestion (6-8 weeks) Triglycerides (mg / dl) Total cholesterol (mg / dl) LDL cholesterol (mg / dl) HDL cholesterol (mg / dl) Arteriosclerosis indices Total cholesterol / HDL cholesterol LDL cholesterol / HDL cholesterol Apolipoprotein A1 (mg / dl) Apolipoprotein B (mg / dl) 215.1 ± 45.5239.6 ± 10.9153.3 ± 10.351.8 ± 3.333.75 ± 0.244.75 ± 0.243.03 ± 0.23143.3 ± 6.0199.9 ± 5.14 195.4 ± 49.9211.8 ± 9.15130.9 ± 8.2658.5 ± 3.383.37 ± 0.244.37 ± 0.242.69 ± 0.20140.2 ± 5.6799.1 ± 5.47

Atherosclerosis index = (total cholesterol-HDL cholesterol) / HDL cholesterol

The results of vascular endothelial dilatation test of the brachial artery are shown in Table 7 and FIG. 12, and FMD, which is an indicator of endothelial-dependent vasodilator function test, is obtained after ingesting the cardiovascular disease improving composition of the present invention. (Flow-mediated dimension) increased by 12%, and Nitroglycerin mediated dimension (NMD), which represents endothelial-independent vasodilator function, increased by 20%.

Vascular endothelial dilatation test results of the brachial artery N = 39 Before intake After ingestion (6-8 weeks) FMD (%) NMD (%) 7.39 ± 0.7010.5 ± 1.08 8.28 ± 1.1912.4 ± 1.05

Example 14

PT, Blood Coagulation Factor Activity and Fibrinogen Concentration after Ingestion of Cardiovascular Disease Improvement Composition

After taking the cardiovascular disease improving composition according to the present invention, the prothrombin time (PT) was increased by about 18% from 8.9 ± 0.2 seconds to 10.5 ± 0.3 seconds, and the activity of each blood coagulation factor was increased. A significant decrease could be observed (see Table 8). In addition, the concentration of fibrinogen in the blood was all within the normal range (see FIG. 13).

 PT, Fibrinogen Concentration and Blood Coagulation Factor Activity Test Results N = 39 Week 0 8 Weeks PT (s) Fibrinogen (mg / dl) Coagulation Factor Fc II (%) Fc V (%) Fc Ⅶ (%) Fc X (%) 8.93 ± 0.24344.2 ± 11.9 132.5 ± 8.33172.3 ± 17.0262.2 ± 26.0120.7 ± 7.89 10.5 ± 0.28306.6 ± 20.6 113.9 ± 9.02155.3 ± 12.2243.2 ± 27.899.5 ± 9.91

Fc (%) = Factor coagulant activity of blood coagulation factor

Blood lipid concentration is a factor that greatly affects the onset and progression of cardiovascular disease, and there have been many correlations between cardiovascular diseases such as arteriosclerosis and blood cholesterol level. It has been proven that it can be effective in the prevention of cardiovascular diseases by improving. As a result of blood analysis, 32 of the 39 subjects (82%) who participated in the study were determined to have a hyperlipidemic threshold (total cholesterol ≥220 mg / dl, LDL cholesterol ≥) as defined by the National Cholesterol Education Program (NCEP). It was determined to be hyperlipidemia above 130 mg / dl or triglyceride ≧ 160 mg / dl). However, the LDL cholesterol concentration was reduced to 15% and the HDL cholesterol level was increased by about 11% by taking the cardiovascular disease improving composition according to the present invention. It has been shown to improve metabolism effectively and to be effective in preventing cardiovascular diseases.

In addition, as a result of functional evaluation and coagulation factor analysis of vascular endothelial cells, the composition showed a tendency to significantly improve the function of vascular endothelial cells and the activity of blood coagulation factors. Endothelial expansion of blood vessels measured in the brachial artery was assessed by FMD (%) for endothelial independent function test and NMD (%) for endothelial independent function test. And it can be seen to have an excellent effect on the improvement of cardiovascular disease.

As described and demonstrated in detail above, the present invention provides a cardiovascular disease improving composition using a bioactive material derived from seaweed. The composition of the present invention has an excellent ability to prevent LDL oxidation and has excellent blood anticoagulant activity so that it can slowly and simultaneously block the main factors of cardiovascular deterioration. In addition, the level of LDL cholesterol was lowered and the level of HDL cholesterol was lowered, thereby lowering the vascular hardening factor, significantly improving the vasodilation function of the coronary atherosclerosis group, and improving the tendency of excessive coagulation to the normal range. On the other hand, the composition was confirmed that there is no toxicity even if ingested 2g per 1kg body weight daily. Therefore, the composition of the present invention can improve hyperlipidemia, improve excessive blood coagulation tendency, and significantly improve blood vessel dilatation ability, and there is no side effect even when ingested in excess, and it is economical, which can effectively prevent and treat cardiovascular diseases, health It can also be widely used in the manufacture of supplements and special nutritional foods.

1 is a bar graph showing the content of total cholesterol when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

2 is a bar graph showing the content of free cholesterol when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

3 is a bar graph showing the content of cholesterol esters when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

4 is a bar graph showing the HDL-cholesterol content when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

5 is a bar graph showing the LDL-cholesterol content when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

6 is a bar graph showing the composition ratio of HDL / LDL-cholesterol when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

7 is a bar graph showing the arteriosclerosis index when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

8 is a bar graph showing the content of triglycerides when the cardiovascular disease improving composition according to the present invention is administered to a hyperlipidemic rat.

9 is a bar graph showing the degree of LDL oxidation according to the concentration of the cardiovascular disease improving composition according to the present invention.

Figure 10 is a bar graph showing the results of the measurement of aPTT carried out to determine the blood anticoagulant activity of the cardiovascular disease improvement composition according to the present invention.

11 is a bar graph showing the total cholesterol, LDL cholesterol and HDL cholesterol levels when taking the cardiovascular disease improving composition according to the present invention in patients with coronary atherosclerosis and hyperlipidemia.

12 is a bar graph showing FMD and NMD when the cardiovascular disease improving composition according to the present invention is administered to a coronary sclerosis patient and a hyperlipidemic patient.

Figure 13 is a bar graph showing the prothrombin time (PT) and fibrinogen concentration when taking the cardiovascular disease improving composition according to the present invention in patients with coronary atherosclerosis and hyperlipidemia.

Claims (7)

2 to 70% by weight of alginic acid derived from seaweed, 0.5 to 30% by weight of fucoidan, 1 to 50% by weight of laminarin, 2 to 70% by weight of phloroglucinol, and 0.01 to 10% by weight of fucosterol. Cardiovascular disease improvement composition. The composition according to claim 1, wherein the composition contains 20-50% by weight of alginic acid, 2-20% by weight of fucoidan, 5-40% by weight of laminarin, 10-50% by weight of phloroglucinol and 0.05-5% by weight of fucosterol. A composition comprising a. The composition of claim 1 or 2, wherein the alginic acid has a molecular weight of 3,000 or more and an average molecular weight of 15,000 to 150,000. The composition according to claim 1 or 2, wherein the fucoidan has a molecular weight of 5,000 or more, an average molecular weight of 20,000 to 200,000, and a sulfated content of 5 to 45%. The method of claim 1 or 2, wherein the phloroglucinol is selected from the group consisting of extract, dieckol, phlorotannin A, phlolofucopurocol, 7,7 'bieckol, 9,9' bieckol and its glycosides. A composition comprising a. The seaweed according to claim 1 or 2, wherein the seaweed is brown seaweed, mabanban, forked bear, blue seaweed, Ecklonia cava, 톳, kelp, rhubarb, seaweed, stalk, dolphin, round stone, steaming, falcon, independence, stone, spirulina , Starfish. Composition comprising at least one selected from the group consisting of head, hearing, chlorella and gompi. The composition according to claim 1 or 2, wherein the composition is added to medicines, dietary supplements, special nutritional products, beverages and liquors, food additives, confectionery or bread.