KR20090010504A - Compositions for the prevention and treatment of cardiovascular diseases containing sophora flavascens extracts, soluble extracts thereof, fractions thereof or flavonoid compounds isolated therefrom as an active ingredient - Google Patents

Abstract

A composition comprising the extract of Sophora flavescenes, soluble extracts thereof, fractions thereof or flavonoids separated therefrom is provided to prevent and treat the heart circulatory system disease. The composition for preventing and treating the heart circulatory system disease comprises the extract of Sophora flavescenes which is prepared by extracting Sophora flavescenes with water, C1-C4 alcohol or mixed solvents thereof, chloroform soluble extracts thereof, fractions thereof which is prepared by performing silica gel chromatography with 100% chloroform, n-hexane/acetone mixed solvent and chloroform/acetone mixed solvent as moving phase or flavonoids separated therefrom.

Description

Composition for the prevention and treatment of cardiovascular diseases containing Sophora flavascens extracts, soluble extracts according to ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds isolated therefrom as an active ingredient , fractions according or flavonoid compounds isolated therefrom as an active ingredient}

The present invention relates to a composition for the prevention and treatment of cardiac circulatory disorders, comprising a ginseng extract, a soluble extract thereof, a fraction thereof or a flavonoid compound isolated therefrom as an active ingredient.

Recently, vascular disorders such as arteriosclerosis have increased significantly along with the increase in adult disease. Atherosclerosis is a typical vascular disorder. Atherosclerosis is a disease in which the arteries are hardened by genetic factors related to lipid metabolism and environmental factors such as eating habits, smoking, and lack of exercise, and are likely to occur in the cerebral artery or coronary artery. It develops into circulatory diseases such as cerebrovascular diseases. In the case of cerebral arteriosclerosis, headache, dizziness, and mental disorders are indicated and cause cerebral dysfunction. In the case of coronary arteriosclerosis, pain and arrhythmia in the heart are known to cause angina pectoris and myocardial infarction. In addition, this causes high blood pressure, heart disease, cerebral hemorrhage, and atherosclerosis is the leading cause of death in modern society, especially among men in their 50s and 60s.

Plaque formation and vascular rupture of the vascular wall are major factors in the development of myocardial infarction, and the hypothesis about the early onset of atherosclerosis is a chronic inflammatory process for damage to the vessel wall. (response-to-injury hypothesis) '(Nature 1993, 362, 801-809). This is a dysfunctional state in which vascular endothelial cells do not maintain normal homeostasis due to genetic variation, peroxide, hypertension, diabetes, increased plasma homocysteine concentration, or microbial infection.

More specifically, these causes cause low-density lipoprotein (LDL) in the blood to change to highly modified-LDL (HM-LDL) through oxidation, glycemic binding, integration, glycoprotein binding, and the like. They stimulate and cause damage to vascular endothelial cells and smooth muscle. As a result, when the expression of vascular cell adhesion molecule-1 (VCAM-1) of endothelial cells and the release of inflammatory mediators of inflammatory cells are promoted, LDL enters and accumulates under the endothelial cells, LDL and oxidized HM-LDL in turn to induce the influx and activation of immune cells, such as macrophages, T lymphocytes to promote the inflammatory response of the lesion. Next, the lesions are necrotic by the action of the introduced macrophages or lymphocytes released from hydrolysases, inflammatory mediators, growth factors, etc., and monocytes are introduced into the necrotic lesions, smooth muscle migration and differentiation, and fibrous tissues. Iterative process such as the formation of a. Through this process, the lesion tissue develops into a fibrous lesion of complex structure covered with fibres on the necrotic tissue with HM-LDL as the nucleus, and a thrombosis is formed from the developed lesion tissue and the arteries are hardened to show circulatory diseases such as blood flow disorder. Will be.

LDL oxidation is most important as an early cause of atherosclerosis, including atherosclerosis (Circulation, 1995, 91, 2488-2496; Arterioscler. Thromb. Vasc. Biol., 1997, 17, 3338-3346 ). Oxidative stress produced in vivo and in vivo transforms LDL in the blood into oxidized-LDL, which enters the intima through an adhesion molecule. Monocytes are introduced into the oxidized-LDL to form foam cells, producing fat streak, an early lesion of atherosclerosis. Atherosclerotic early lesions are characterized by the expression of VCAM-1, intracellular adhesion molecule-1 (IMC-1) and monocyte chemoattractant protein-1 (MCP-1), which are produced by arterial endothelial cells. It is induced by the transcription factor (NF-κB). In addition, NF-κB causes plaque formation and rupture in the vessel wall. NF-κB is activated by various factors such as reactive oxygen species or cytokines, and is a heterodimer composed of p50 and p65, which is a transcription factor involved in regulating the target gene of many cells. to be. Activated NF-κB binds to specific promoter genes such as IL-1, VCAM-1, ICAM-1, and other factors involved in the progression of atherosclerosis, resulting in the expression of various inflammatory factors. Adjust.

Antioxidants and free radical scavengers, so is found to inhibit this NF-κB activity in case of ingestion of antioxidants as appropriate, in vivo (in In vivo ) is expected to inhibit the arteriosclerosis by inhibiting the oxidation of LDL, inhibit the expression of adhesion molecules and decrease the activity of NF-κB (Korean Patent Publication No. 2003-0014155) . In addition, studies on the factors and removal of LDL peroxide in hyperlipidemia and atherosclerosis patients (Curr. Atheroscler. Res., 2000, 2, 363-372) are also actively conducted.

In addition, high blood cholesterol levels are likely to cause coronary cardiovascular disease. Therefore, in order to reduce blood cholesterol levels, it is necessary to suppress the absorption of cholesterol by dieting to reduce the intake of cholesterol and fat or by inhibiting enzymes related to lipid metabolism. . Accordingly, many studies have been conducted on acyl-CoA (cholesterol acyltransferase; ACAT), an enzyme that esterifies cholesterol.

The mechanism of action of ACAT occurs largely in three parts of the intestinal, liver and vascular wall cells of the body.

First, in the intestine, ACAT converts ingested cholesterol into the form of esters to facilitate its absorption into the intestine. Second, cholesterol that is absorbed from the outside or biosynthesized in the body is accumulated in a carrier called very low-density lipoprotein (VLDL) in the liver and then supplied to each organ of the body through blood vessels. The conversion allows for the accumulation of cholesterol in the carrier. Third, in the cells that make up the arterial vessel wall, ACAT converts cholesterol into its ester form to promote the accumulation of cholesterol in the cell, which is a direct cause of atherosclerosis. In addition, since foam cells contain a large amount of cholesteryl ester derived from cholesterol by ACAT activity, the formation of foam cells derived from macrophages and smooth muscle cells is very important from an experimental and clinical aspect. Since the proliferation of foam cells in the vessel wall is directly related to increased ACAT activity, the development of ACAT inhibitors as a potent antiarterial agent is desirable.

Therefore, the drug that inhibits the activity of ACAT may first reduce the amount of cholesterol that enters the body by inhibiting the absorption of intestinal cholesterol, and secondly, by inhibiting the release of cholesterol into the blood vessels from the liver to reduce blood cholesterol levels. Third, it is expected to prevent arteriosclerosis by preventing cholesterol from accumulating in blood vessel wall cells.

ACAT activity inhibitors reported to date are activity inhibitors for rat hepatic microsomal ACAT or rat hepatic macrophage (J774) ACAT. Human ACAT includes hACAT-1 (50 kDa) and hACAT-2 (46 kDa), hACAT-1 mainly acts on the liver, adrenal glands, macrophages and kidneys of adults, while hACAT-2 acts on the small intestine and liver Rudel, LL et al ., Curr. Opin. Lipidol. 12, 121-127, 2001). Substances that inhibit human ACAT activity are targets for the prevention and treatment of hypercholesterolemia, cholesterol stones or atherosclerosis through mechanisms that inhibit the absorption of cholesterol from food and the accumulation of cholesteryl esters in the blood vessel walls. (Buhman, KK et al ., Nature Med. 6, 1341-1347, 2000).

Probucol, N, N'-diphenylenediamine (N, N'-diphenylenediamine), phenolic synthetic antioxidants BHA (butylated hydroxyanisol) and BHT (butylated hydroxytoluene), which are currently used as a treatment for hyperlipidemia, are cholesterol in LDL. And reduce the lesion by reducing the degree of oxidation, the antihyperlipidemic agent is excellent in antioxidant power, but its use is limited because of its many side effects.

Thus, attempts have been made to reduce plasma LDL levels through inhibition of cholesterol absorption and inhibition of biosynthesis (Principles in Biochemistry, lipid biosynthesis, 770-817, 3rd Edition, 2000 Worth Publishers). , New York; Steinberg, N. Engl. J. Med., 1989, 320, 915-924). In addition, in patients with hyperlipidemia or atherosclerosis, there is a growing interest in the concurrent administration of lipid-lowering agents with LDL antioxidants.

Meanwhile, Sophora flavescens is a perennial herb that is distributed in Korea, Japan, China, and Siberia. In oriental medicine, dried roots are called ginseng and are used for flavoring, indigestion, neuralgia, hepatitis, jaundice, and hemorrhoids. In private, stems and leaves are used as antipyretic and analgesic drugs. Korean ginseng has been studied in Japan since 1957, and alkaloids (Saito, K. et. al ., Planta Med ., 56: 487, 1990; Atta. R. et al ., J. Nat. Prod ., 54: 929, 1991) and flavonoids (Iinuma, M. et. al ., Phytochemistry , 29, 2667, 1990; Shirataki, Y. et al ., Chem . Pharm . Bull . , 38, 1712, 1990). Especially, in ginseng, it is well known that flavonoids contain a large amount of flavonoids substituted with prenyl groups having 5 carbon isoprenyl groups and 10 carbon lavendulyl groups in the flavonoid structure (Kim, SJ et. al ., Biol . Pharm . Bull ., 26, 1348, 2003) However, activity studies on these have been shown to be cytotoxic (Kang, TH et al ., J. Nat . Prod ., 63, 680, 2000) and anti-inflammatory (Chi, YS et al ., Biochem . Pharm ., 62, 1185, 2001) and its effects on hyperlipidemia and atherosclerosis-related metabolism have not been published.

Therefore, the present inventors, while searching for a new hyperlipidemia or arteriosclerosis treatment agent with fewer side effects in natural products, the antioxidant activity against LDL and inhibitory effect against LCAT in the ginseng extract, its soluble extract, its fractions or flavonoid compounds isolated therefrom Confirmed that the present invention was completed.

It is an object of the present invention to provide a composition for the prevention and treatment of cardiovascular disease, which contains a ginseng extract, a soluble extract thereof, a fraction thereof or a flavonoid compound isolated therefrom as an active ingredient.

In order to achieve the above object, the present invention provides a composition for the prevention and treatment of heart circulatory diseases, containing a ginseng extract, a soluble extract thereof, a fraction thereof or a flavonoid compound isolated therefrom as an active ingredient.

Hereinafter, the present invention will be described in detail.

The ginseng extract of the composition according to the present invention is preferably obtained by extracting from the root of the ginseng, the ginseng can be used without limitation, such as those grown or commercially available. The method of preparing the ginseng extract may use a conventional extraction method in the art. The ginseng extract can be obtained by extracting dried ginseng with water, C ₁ ~ C ₄ alcohol, or a mixed solvent thereof, preferably with methanol, ethanol or butanol. In particular, it is more preferable to extract with ethanol. At this time, the amount of the extraction solvent is preferably 2 to 20 times the dry weight of the ginseng. In the case of using water, C ₁ ~ C ₄ alcohol, or a mixed solvent thereof as the extraction solvent, it can be extracted by standing for 2 to 20 days at room temperature. In addition, the extract may be prepared by extracting ginseng with hot water. When extracted with hot water, the dried ginseng dried material can be extracted by boiling with distilled water and left for 1 to 3 hours. The extraction process may be repeated several times, after which it may be further subjected to methods such as filtration, purification, concentration or lyophilization.

Soluble extract of the composition according to the present invention is concentrated by removing the solvent to remove the ginseng extract, and then suspended in water, alcohol of C ₁ ~ C ₄ , or a mixed solvent thereof, with n-hexane, chloroform and ethyl acetate N-hexane soluble extract, chloroform soluble extract or ethyl acetate soluble extract obtained by sequential fractionation. Chloroform soluble extract is preferred among the soluble extracts. This process may use a conventional fractional extraction method, preferably a fractional funnel. At this time, the suspension solvent is preferably a mixed solvent of water and methanol, the mixed solvent is preferably water: methanol is mixed in a volume ratio of 9: 1.

Fractions of the composition according to the present invention is 100% chloroform, n-hexane / acetone (100: 1 ~ 1: 5, volume ratio) mixed solvent and chloroform / acetone (150: 1 ~ 1: 4, relative to the chloroform soluble extract Volume ratio) can be obtained by performing a silica gel chromatography with a mixed solvent as the mobile phase. At this time, the silica gel is preferably 230 ~ 400 mesh (mesh).

A specific fraction obtained by performing the silica gel chromatography was used as a mobile phase by using a chloroform / acetone (80: 1 to 1: 4, volume ratio) mixed solvent or n-hexane / acetone (100: 1 to 1: 1, volume ratio) mixed solvent. Six flavonoid compounds can be obtained by using the resultant, and performing further silica gel chromatography under reduced pressure and concentration.

Preferred examples of the flavonoid compound are as follows.

1) 5,7,2 ', 4'-tetrahydroxy-8-rabbendulylflavanone;

2) 5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone;

3) 5,7,4'-trihydroxy-8-rabbendulyl-2'-methoxyflavanone;

4) 5,7,2'-trihydroxy-8-rabbendulylflavanone;

5) 5,7,2'-trihydroxy-8- (5-hydroxy-2-isopropenyl-5-methylhexyl) -flavanone; or

6) 5,2'-dimethoxy-7,4'-dihydroxy-8-rabbendulylflavanone.

The chromatography may be performed once or several times until a single compound is purified, and may be concentrated and recrystallized as necessary. The separated compound can be analyzed by conventional spectroscopic analytical methods such as molecular weight measurement and NMR measurement.

Structural formulas of the six compounds obtained according to the above method are shown in Table 1 below.

Chemical formula rescue denomination One

5,7,2 ', 4'-tetrahydroxy-8-lavendulylflavanone (Sophoraflavanone G) 2

5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone ((-)-curanone) 3

5,7,4'-Trihydroxy-8-lavendulyl-2'-methoxyflavanone (lysianone A) 4

5,7,2'-trihydroxy-8-lavendrilylflavanone (Cusenol A) 5

5,7,2'-trihydroxy-8- (5-hydroxy-2-isopropenyl-5-methylhexyl) -flavanone (cusenol T) 6

5,2'-dimethoxy-7,4'-dihydroxy-8-rabbendulylflavanone ((2S) -2'-methoxy-curanone)

The flavonoid compound may be used in the form of a pharmaceutically acceptable salt, and all salts, hydrates and solvates prepared by conventional methods, commercially available reagents may be used, and in the present invention, extraction, separation, and purification from red ginseng Use it.

The ginseng extract, the soluble extract thereof, the fraction thereof or the flavonoid compound isolated from the ginseng extract contained in the composition according to the present invention is characterized by having antioxidant activity and acyl coei: cholesterol acyl transferase inhibitory activity.

In order to confirm the antioxidant activity of the ginseng extract, soluble extract thereof, fraction thereof or flavonoid compound isolated from the composition according to the present invention, the antioxidant activity against LDL was determined by TBARS (thiobarbituric acid-reactive substances) method. As a result of the measurement, the extract of red ginseng, its soluble extract, its fraction or its flavonoid compounds isolated from it was found to have excellent antioxidant activity against LDL. In particular, 5,7,2 ', 4'-tetrahydroxy-8-lavendrilyl flavanone and 5-methoxy-7,2', 4'-trihydroxy-8-rabbendulyl flavanone which are flavonoid compounds Showed excellent oxidation inhibition of at least 80% (see Table 2 ).

In addition, to determine the acyl coay: cholesterol acyl transferase inhibitory activity of the ginseng extract, soluble extract thereof, fraction thereof or flavonoid compound isolated therefrom according to the present invention, human ACAT-1 and ACAT As a result of the experiment performed on -2, the ginseng extract, its soluble extract, its fraction or the flavonoid compound isolated therefrom was found to be excellent in inhibiting ACAT activity. In particular, the flavonoid compound 5,7,2'-trihydroxy-8-rabbendulylflavanone inhibited the activity of human ACAT-1 by 70% or more, and inhibited the activity of human ACAT-2 by 50% or more ( See Table 3 ).

ACAT activity inhibitors reported to date are activity inhibitors for rat hepatic microsomal ACAT or rat hepatic macrophage (J774) ACAT. Human ACAT includes hACAT-1 (50 kDa) and hACAT-2 (46 kDa), hACAT-1 mainly acts on the liver, adrenal glands, macrophages and kidneys of adults, while hACAT-2 acts on the small intestine and liver Rudel, LL et al ., Curr. Opin. Lipidol. 12, 121-127, 2001). Substances that inhibit human ACAT activity are targets for the prevention and treatment of hypercholesterolemia, cholesterol stones or atherosclerosis through mechanisms that inhibit the absorption of cholesterol from food and the accumulation of cholesteryl esters in the blood vessel walls. (Buhman, KK et al ., Nature Med. 6, 1341-1347, 2000). Probucol, N, N'-diphenylenediamine (N, N'-diphenylenediamine), phenolic synthetic antioxidants BHA (butylated hydroxyanisol) and BHT (butylated hydroxytoluene), which are currently used as a treatment for hyperlipidemia, are cholesterol in LDL. It is reduced, the degree of oxidation is reduced to reduce the lesion formation, the antihyperlipidemic agent is excellent in antioxidant power, but its use is limited because of many side effects.

Attempts have been made to reduce plasma LDL levels through inhibition of cholesterol absorption and inhibition of biosynthesis (Principles in Biochemistry, lipid biosynthesis, 770-817, 3rd Edition, 2000 Worth Publishers, New). York; Steinberg, N. Engl. J. Med., 1989, 320, 915-924). In addition, in patients with hyperlipidemia or atherosclerosis, there is a growing interest in the concurrent administration of lipid-lowering agents with LDL antioxidants.

Therefore, the ginseng extract, soluble extract thereof, fractions thereof, or flavonoid compounds isolated therefrom according to the present invention have antioxidant activity and acyl coei: cholesterol acyl transferase inhibitory activity, and thus, the composition of the present invention. May be usefully used as a prophylactic and therapeutic agent for cardiovascular diseases such as hyperlipidemia, coronary heart disease, arteriosclerosis, and myocardial infarction, which are known to be induced by the oxidation of LDL or the synthesis and accumulation of cholesteryl esters.

The composition of the present invention may contain one or more active ingredients exhibiting the same or similar functions in addition to the ginseng extract, soluble extract thereof, fractions thereof, or flavonoid compounds isolated therefrom.

The composition of the present invention may be prepared by including one or more pharmaceutically acceptable carriers in addition to the active ingredient described above for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, as an antioxidant, buffer And other conventional additives such as bacteriostatic agents. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by an appropriate method in the art or using a method disclosed in Remington's Pharmaceutical Sciences (Mack Publishing Company, Easton PA).

The compositions of the present invention may be administered parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) or orally, depending on the desired method, and the dosage may be based on the weight, age, sex, health status, The range varies depending on the diet, the time of administration, the method of administration, the rate of excretion and the severity of the disease. The daily dosage is 0.1-1000 mg / kg, preferably 0.1-500 mg / kg, based on the ginseng extract, soluble extract thereof or fractions thereof contained in the composition of the present invention. More preferred. The daily dose of the flavonoid compound is 0.1 to 100 mg / kg, preferably 0.5 to 10 mg / kg, and more preferably administered once to several times a day.

As a result of the oral administration of mouse ginseng extract, soluble extract thereof, fraction thereof or flavonoid compound isolated from the composition according to the present invention to mice, the ginseng extract, soluble extract thereof, fraction thereof or Flavonoid compounds were found to have a 50% lethal dose (LD ₅₀ ) of at least 1000 mg / kg or more by oral toxicity test (see Experimental Example 3).

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

In addition, the composition according to the present invention can be added to health food for the purpose of improving cardiovascular disease. When the ginseng extract, its soluble extract, its fraction or flavonoid compound separated therefrom is used as a food additive, the ginseng extract, its soluble extract, its fraction or flavonoid compound separated therefrom is added as is or other food or It can be used together with food ingredients and can be suitably used according to conventional methods. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, in the manufacture of food or beverages, the ginseng extract, its soluble extract, its fractions or flavonoid compounds separated therefrom are added in an amount of 1 to 20% by weight, preferably 5 to 10% by weight based on the raw materials. do. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

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

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The natural carbohydrates are sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

The composition of the present invention can be used in various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents and the like used in beverages.

In addition, the composition of the present invention may contain a flesh for the production of natural fruit juice, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical, but the composition of the present invention is generally selected in the range of 0 to 20 parts by weight per 100 parts by weight.

As described above, the ginseng extract, the soluble extract thereof, the fraction thereof or the flavonoid compound separated therefrom, which are included in the composition according to the present invention, have very high antioxidant activity against low-density lipoprotein (LDL). In addition to being excellent, it effectively inhibits the activity of acyl-CoA (cholesterol acyltransferase (ACAT), thus the inventive preparations can be used for the synthesis and accumulation of LDL oxidation or cholesteryl esters. It can be usefully used as a prophylactic and therapeutic agent for cardiovascular diseases such as hyperlipidemia, coronary heart disease, arteriosclerosis, and myocardial infarction which are known to be induced.

Hereinafter, the present invention will be described in detail through examples. These examples are for illustrating the present invention in detail, and the scope of the present invention is not limited by these examples.

< Example  1> Gosam  Preparation of Extract

<1-1> Gosam  Preparation of Ethanol Extract

The ginseng used in this experiment was purchased from Chinese herbal medicines in Jinju, Gyeongsangnam-do. 1 kg of purchased ginseng was added to 10 l of 95% ethanol, and extracted two times while standing at room temperature for 10 days. The extract was filtered through filter paper (Advantec N. 2, filter paper) to remove the precipitate and afford a filtrate. The filtrate was concentrated under a reduced pressure to obtain an ethanol extract (44 g) as a brown oily substance.

<1-2> Gosam Hydrothermal  Extract manufacturer

To 100 g of ginseng purchased on the medicinal herb, 1 L of distilled water was added, followed by extraction for 2 hours with stirring. The extraction process was repeated twice. The extract obtained above was filtered through filter paper (Advantec N0.2, filter paper) to remove precipitates and to obtain a filtrate. The filtrate was concentrated under reduced pressure to obtain a ginseng hot water extract (14 g).

< Example  2> Gosam  Isolation and Purification of Flavonoid Compounds from Extracts

The ginseng ethanol extract (44 g) obtained in Example 1 was suspended in 2.0 L of a mixed solution of water and methanol (9: 1 volume ratio), and then partitioned into n-hexane and chloroform in order to dissolve 14.3 g of n -hexane. The oily substance of the extract and 29.8 g of soluble chloroform extract was obtained.

<2-1> 5,7,2 ', 4'- Tetrahydroxy -8- Ravendullil Flavanon  detach

Silica gel column chromatography (silica gel 450 g. 230-400 mesh) using a mixed solvent of 100% chloroform and chloroform / acetone (150: 1 to 1: 2 volume ratio) as a mobile phase with respect to 29.8 g of the chloroform soluble extract obtained above. ) Was separated into 35 fractions (Fr.1-35, 30 ml / fraction). For the sixth fraction (Fr. 6, 1.2 g), silica gel column chromatography (200 g, 230-400 mesh) was again performed by using a mixed solvent of chloroform / acetone (40: 1 to 10: 1 by volume) as a mobile phase. Twenty fractions (Fr. 6-1 to 6-20, 20 ml / fraction) were obtained. The 11 to 15 fractions (Fr. 6-11 to 6-15) were concentrated under reduced pressure and concentrated to give 5,7,2 ', 4'-tetrahydroxy-8-rabbendulylflavanone as a yellow needle (1). , 110 mg) was obtained.

m.p. 178 to 180 ° C;

Molecular weight 424;

Molecular formula C ₂₅ H ₂₈ O ₆ ;

¹ H-NMR (MeOD, 500 MHz) δ 1.48 (3H, s, H-6 "), 1.56 (3H, s, H-7"), 1.63 (3H, s, H-10 "), 2.00 (2H , m, H-3 "), 2.47 (1H, m, H-2"), 2.58 (2H, m, H-1 '), 2.73 (1H, dd, J = 17.1, 2.8 Hz, H-3a) , 2.97 (1H, dd, J = 17.1, 13.2 Hz, H-3b), 4.55 (2H, m, H-9 "), 4.90 (1H, m, H-4"), 5.56 (1H, dd, J = 13.2, 2.7 Hz, H-2), 5.92 (1H, s, H-6), 6.35 (1H, m, H-5 '), 6.37 (1H, d, J = 2.3 Hz, H-3') , 7.30 (1H, doublet, J = 8.2 Hz, H-6 ');

EIMS m / z 424 [M] ⁺ (10), 301 (80), 283 (100), 219 (13), 165 (80).

<2-2> 5- Methoxy -7,2 ', 4'- Trihydroxy -8- Ravendullil Flavanon  detach

Silica gel column chromatography (180 g, 230-400 mesh) was carried out again using the mixed solvent of hexane / acetone (100: 1 to 1: 1 by volume) for the eighth fraction (Fr.8, 1.3 g) of the chloroform soluble extract. ) To obtain 40 fractions (Fr. 8-1 to 8-40, 10 ml / fraction). The 22 to 26 fractions (Fr.8-22 to 8-26) were concentrated under reduced pressure and concentrated to give an orange needle of 5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone (2 , 50 mg) was obtained.

m.p. 117 to 121 ° C;

Molecular weight 438;

Molecular formula C ₂₆ H ₃₀ O ₆ ;

¹ H-NMR (MeOD, 500 MHz) δ 1.46 (3H, s, H-6 ″), 1.56 (3H, s, H-7 ″), 1.63 (3H, s, H-10 ″), 1.99 (2H , m, H-3 "), 2.49 (1H, m, H-2"), 2.62 (2H, m, H-1 "), 2.69 (1H, dd, J = 16.7, 2.8 Hz, H-3b) , 2.87 (1H, doublet, J = 16.7, 13.2 Hz, H-3a), 3.80 (3H, s, -OCH ₃ ), 4.51 (1H, m, H-9 "b), 4.57 (1H, m, H -9 "a), 4.96 (1H, m, H-1"), 5.54 (1H, dd, J = 13.2, 2.7 Hz, H-2), 6.10 (1H, s, H-6), 6.34 (1H , m, H-5 '), 6.36 (1H, d, J = 2.3 Hz, H-3'), 7.29 (1H, d, J = 8.2 Hz, H-6 ');

EIMS m / z 438 [M] ⁺ (5), 422 (18), 418 (18), 299 (100), 179 (28), 153 (32).

<2-3> 5,7,4'- Trihydroxy -8- Ravendullil -2'- Methoxyflavanone  detach

Silica gel column chromatography (230 g, 230-400) with a mixed solvent of chloroform / acetone (80: 1 to 10: 1 by volume) for the thirteenth fraction (Fr. 13, 1.5 g) of the chloroform soluble extract was used as a mobile phase. Mesh) to obtain 50 fractions (Fr. 13-1 to 13-50, 10 ml / fraction). The 31 to 38 fractions (Fr. 13-31 to 13-38) were concentrated under reduced pressure and concentrated to give a yellow needle-like 5,7,4'-trihydroxy-8-lavendilyl-2'-methoxyflavanone (3 , 70 mg) was obtained.

m.p. 155 to 156 ° C;

Molecular weight 438;

Molecular formula C ₂₆ H ₃₀ O ₆ ;

¹ H-NMR (Acetone- d ₆ , 500 MHz) δ 1.49 (3H, s, 7 "-Me), 1.57 (3H, s, 6" -Me), 1.64 (3H, s, 10 "-Me), 2.08 (2H, m, H-3 "), 2.53 (1H, m, H-2 '), 2.65 (2H, m, H-1"), 2.72 (1H, dd, J = 16.8, 2.9 Hz, H -3), 3.03 (1H, dd, J = 16.8, 13.2 Hz, H-3), 3.83 (3H, s, OMe), 4.55 (2H, m, H-9 "), 2.97 (1H, t, J = 6.9 Hz, H-4 "), 5.68 (1H, dd, J = 13.2, 2.9 Hz, H-2), 6.01 (1H, s, H-6), 6.51-6.57 (2H, m, H-3 ', H-5'), 7.42 (1H, d, J = 8.1 Hz, H-6 '), 8.52 (1H, br s, 4'-OH), 9.91 (1H, br s, 7-OH), 12.16 (1H, s, C ₅ -OH);

EIMS m / z 438 [M] ⁺ (15), 423 (12), 315 (100), 219 (11), 165 (80), 135 (10).

<2-4> 5,7,2'- Trihydroxy -8- Ravendullil Flavanon  detach

Silica gel column chromatography (50 g, 230-400) was carried out using the mixed solvent of chloroform / acetone (50: 1 to 5: 1 by volume) as the mobile phase for the fifteenth fraction (Fr. 15, 300 mg) of the chloroform soluble extract. Mesh) to give 15 fractions (Fr. 15-1 to 15-15, 10 ml / fraction). The 7-10 fraction (Fr. 15-7-15-15) was concentrated under reduced pressure, and the white powder 5,7,2'- trihydroxy-8- rabendulyl flavanone (4, 80 mg) was obtained.

m.p. 175 to 176 ° C .;

Molecular weight 408;

Molecular formula C ₂₅ H ₂₈ O ₅ ;

¹ H-NMR (CDCl ₃ , 500 MHz) δ 1.46 (3H, s, H-6 ″), 1.55 (3H, s, H-7 ″), 1.64 (3H, s, H-9 ″), 2.00 ( 2H, m, H-3 "), 2.48 (1H, m, H-2"), 2.61 (2H, m, H-1 "), 2.87 (2H, m, H-3), 4.59 (2H, m , H-10 "), 4.96 (1H, m, H-4"), 5.63 (1H, dd, J = 12.24, 3.7 Hz, H-2), 5.93 (1H, s, H-6), 6.83 ( 1H, dd, J = 8.12, 0.9 Hz, H-3 '), 6.90 (1H, ddd, J = 15.04, 7.6, 0.9 Hz, H-5'), 7.17 (1H, ddd, J = 15.47, 7.9, 1.6 Hz, H-4 '), 7.54 (1H, doublet of doublets, J = 7.6, 1.4 Hz, H-6');

EIMS m / z 408 [M] ⁺ (100), 286 (100), 268 (100), 220 (40), 120 (70).

<2-5> 5,7,2'- Trihydroxy -8- (5-hydroxy-2- Isopropenyl -5- Methylhexyl )- Flavanese  detach

Silica gel column chromatography (80 g, 230-400 mesh) was carried out using the mixed solvent of chloroform / acetone (40: 1 to 2: 1 volume ratio) as a mobile phase for the twentieth fraction of the chloroform soluble extract (Fr.20, 800 mg). ) To obtain 30 fractions (Fr. 20-1 ~ 20-30). The 21 to 23 fractions (Fr. 20-21 to 20-23, 5 ml / fraction) were concentrated under reduced pressure and concentrated to give a yellow needle-like 5,7,2'-trihydroxy-8- (5-hydroxy-2 -Isopropenyl-5-methylhexyl) -flavanone (5, 30 mg) was obtained.

m.p. 165 to 166 ° C .;

Molecular weight 426;

Molecular formula C ₂₅ H ₃₀ O ₆ ;

¹ H-NMR (CDCl ₃ , 500 MHz) δ 1.09 (3H, s, H-6 ″), 1.16 (3H, s, H-7 ″), 1.63 (3H, s, H-10 ″), 1.25- 1.50 (4H, m, H-3 ", 4"), 2.20 (1H, dd, J = 13.6, 5.6 Hz, H-1 "), 2.62 (1H, dd, J = 13.6, 8.4 Hz H-1" ), 2.89 (1H, dd, J = 17.6, 2.4 Hz, H-3a), 3.03 (1H, dd, J = 17.6, 13.2 Hz, H-3b), 4.62 (1H, s, H-9 "), 4.69 (1H, s, H-9 "), 5.67 (1H, dd, J = 13.2, 2.4 Hz, H-2), 6.02 (1H, s, H-6), 6.87 (1H, d, J = 8.4 Hz, H-3 '), 6.98 (1H, t, J = 7.6 Hz, H-5'), 7.22 (1H, d, J = 8.4 Hz, H-6 '), 12.18 (1H, s, OH C -5);

EIMS m / z 426 [M] ⁺ (5), 408 (10), 285 (100), 267 (80), 165 (65).

<2-6> 5,2'- Dimethoxy -7,4'-dihydroxy-8- Ravendullil Flavanon  detach

Silica gel column chromatography (50 g, 230-400 mesh) was carried out using a mixed solvent of chloroform / acetone (10: 1 to 1: 4 by volume) for the thirty second fraction of the chloroform soluble extract (Fr.32, 400 mg). 35 fractions (Fr. 32-1 to 32-35, 5 ml / fraction) were obtained. The 11 to 16 fractions (Fr. 32-11 to 32-16) were concentrated under reduced pressure and concentrated to give a yellow needle-like 5,2'-dimethoxy-7,4'-dihydroxy-8-rabbendulylflavanone (6 , 20 mg) was obtained.

m.p. 112 to 115 ° C;

Molecular weight 452;

Molecular formula C ₂₇ H ₃₂ O ₆ ;

¹ H-NMR (CDCl ₃ , 500 MHz) δ 1.45 (3H, s, H-6 ″), 1.56 (3H, s, H-7 ″), 1.63 (3H, s, H-9 ″), 2.00 ( 2H, m, H-3 "), 2.49 (1H, m, H-2"), 2.62 (2H, m, H-1 "), 2.69 (1H, dd, J = 16.7, 2.9 Hz, H-3 ), 2.87 (1H, dd, J = 16.7, 13.2 Hz, H-3), 3.79 (3H, s, H-11 "), 3.80 (3H, s, H-12"), 4.57 (2H, m, H-10), 4.96 (1H, m, H-4 "), 5.54 (1H, dd, J = 13.2, 2.7 Hz, H-2), 6.10 (1H, s, H-6), 6.34 (1H, s, H-5 '), 6.36 (1H, d, J = 2.3 Hz, H-3'), 7.34 (1H, d, J = 8.2 Hz, H-6 ');

EIMS m / z 452 [M] ⁺ (10), 329 (25), 233 (95), 179 (100).

Experimental Example 1 Measurement of Antioxidant Activity of Red Ginseng Extract, its Soluble Extract, Fractions thereof, or Flavonoid Compounds Separated from

In order to determine the antioxidant activity against the LDL of the ginseng extract, its soluble extract, its fraction or the flavonoid compound isolated therefrom, the experiment was carried out as follows.

Cu ^{2 +} is known to induce the oxidation of LDL. Therefore, in the present invention, Cu ²⁺ is a thiobarbituric acid-reactive substances (TBARS) method of dialdehyde (dialdehyde) which is an oxidation product of unsaturated fatty acids produced by inducing oxidation of LDL, and included in the composition according to the present invention. The antioxidant activity of the ginseng extract, soluble extract thereof, fraction thereof or flavonoid compound isolated therefrom was investigated (Packer, L. Ed. (1994) Methods in Enzymology Vol. 234, Oxygen radicals in biological systems Part D. Academic press, San Diego).

1-1. LDL Separation of

LDL was isolated with a slight modification of the standard method. First, whole blood was collected from healthy healthy volunteers through the Korean Red Cross Blood Source. The blood was centrifuged at 4 ° C. at 3000 revolutions per minute (rpm) for 30 minutes to obtain plasma, followed by EDTA (0.1%), NaN ₃ (0.05%), to prevent denaturation of lipoproteins. PMSF (0.015%) was added and centrifuged at 43800 rpm (100000 g) in a Beckman T8-M ultracentrifuge for 20 hours. The upper layer suspended chylomicron and very low-density lipoprotein (VLDL) were removed, and the remaining solution was collected, and then the specific gravity of the solution was adjusted to 1.063 g / ml using NaBr solution. Thereafter, LDL (1.006 <d <1.063 g / ml) in the upper portion was separated by centrifugation at 43800 rpm for 20 hours at 4 ° C., and dialyzed at 4 ° C. using 10 mM PBS (pH 7.4). . The dialyzed LDL was used within 4 weeks after storage at 4 ° C. Purity of LDL was confirmed by agarose gel electrophoresis and SDS-PAGE (Chapman MJ, et al., J. Lipid Res. 22: 339-358, 1981), and the content of LDL protein was determined by BSA (bovine serum albumin). Quantification was made as a standard (Mahley, RW, et al., J. Lipid Res. 1984; 25: 1277-1294).

1-2. TBARS ( thiobarbituric acid reactive substances ) Measure

20 μl of the isolated LDL (protein concentration, 50-100 μg / ml) was mixed with 210 μl of 10 mM phosphate-buffered saline (PBS), and the red ginseng extract prepared in the above example was used. 10 μl each of an extract, a fraction thereof, or a solution of the flavonoid compound separated therefrom was added.

Red ginseng extract, its soluble extract, its fractions or flavonoid compounds isolated therefrom were dissolved in DMSO (dimethylsulfoxide) and diluted to different concentrations before use in experiments. As the negative control, only the solvent was added.

10 μl of 0.25 mM copper sulfate was added to the solution for 4 hours at 37 ° C., and 1 ml of 20% trichloroacetic acid (TCA) solution was added to stop the reaction. 1 ml of a 0.67% TBA solution dissolved in 0.05 N sodium hydroxide solution was added, stirred for 10 seconds, and heated at 95 ° C. for 5 minutes to allow a color reaction to occur, and the solution was cooled with ice water. The solution was centrifuged at 3000 rpm for 5 minutes to separate the supernatant, and the absorbance at 540 nm was measured by UV-Vis spectroscopy to determine the amount of malondialdehyde (MDA) produced by the color reaction. .

Using a stock solution of tetramethoxypropane (malonaldehyde bis (dimethylacetal)), malondialdehyde was diluted to 0-10 nmol with a phosphate buffer solution to prepare a standard. 250 μl of this standard was developed in the same manner as described above to determine the absorbance at 540 nm and to obtain the standard curve of malondialdehyde Ginseng extract, soluble extract thereof, fractions thereof or flavonoids isolated therefrom. The amount of malondialdehyde in the experiment using the compound was quantified using this standard curve, the results are shown in Table 2 below.

sample Extract or compound LDL oxidation inhibition rate (%, 10 μg / ml) Ginseng extract Ethanol extract 88.5% inhibition at 40 μg / ml Hydrothermal extract 48.1% inhibition at 40 μg / ml n-hexane soluble extract 46.5% inhibition at 40 μg / ml Chloroform Soluble Extract 97.0% inhibition at 40 μg / ml Ethyl acetate soluble extract 99.0% inhibition at 40 μg / ml Flavonoid Compounds 5,7,2 ', 4'-tetrahydroxy-8-rabbendulylflavanone 83.3 5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone 84.3 5,7,4'-trihydroxy-8-lavendulyl-2'-methoxyflavanone 22.5 5,7,2'-trihydroxy-8-lavendulylflavanone 37.8 5,7,2'-trihydroxy-8- (5-hydroxy-2-isopropenyl-5-methylhexyl) -flavanone 31.4 5,2'-dimethoxy-7,4'-dihydroxy-8-lavendulylflavanone 10.5

As shown in Table 2 , the ginseng extract, soluble extract thereof, fractions thereof, or flavonoid compounds separated therefrom contained in the composition according to the present invention were excellent in antioxidant activity against LDL. In the case of red ginseng extract, ethanol extract, and soluble extract, chloroform soluble extract and ethyl acetate soluble extract showed more than 85% inhibition at 40 ㎍ / ml, especially the flavonoid compound 5,7,2 ', 4'-tetra Hydroxy-8-rabbendulylflavanone and 5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone showed excellent oxidation inhibition of at least 80%.

Experimental Example 2 Effect of Ginseng Extract, Soluble Extract, Fractions, and Flavonoid Compounds in ACAT Activity in the Composition

To examine the effect on the ACAT activity of the ginseng extract, its soluble extract, its fraction and flavonoid compounds of the composition according to the present invention, the following experiment was performed.

2-1. ACAT Enzyme  Produce

In order to determine the effect on the activity of human ACAT-1 and ACAT-2, the proteins of hACAT-1 and hACAT-2 were obtained using a baculovirus expression system. Viruses were prepared by inserting cDNAs of hACAT-1 and hACAT-2, respectively, obtained through human liver cDNA library screening into baculovirus delivery vectors, and introducing into insect cells sf9 cells. Next, after separating the recombinant viruses of each of the hACAT-1 and hACAT-2 by plaque purification method, the titer of the storage virus (viral stock) was increased through three amplification processes. Recombinant virus was infected with Hi5 insect cells having good protein expression efficiency so as to have a multiplicity of infection (Mutiplicity of Infection) of 1, followed by shaking culture at 27 ° C for one day. In order to separate microsomal fractions from Hi5 cells overexpressed in cultured hACAT-1 and hACAT-2, the cells were collected by centrifugation at 500 rpm for 15 minutes and rapidly frozen and thawed in a hypotonic buffer. After breaking the cells with ultracentrifugation for one hour at 100000 rpm. The obtained microsome fractions were suspended in storage buffer so that the protein concentration was 8 mg / ml and stored in a cryocooler until use.

2-2. ACAT  Active measurement

Determination of ACAT activity was carried out by the method of Brecher & Chan (P. Brecher and C. Chan, Biochim. Biophys ) using [1- ¹⁴ C] oleoyl-CoA (56.0 μCi / μmol; Amersham) as substrate . a. Acta, 617, 458, 1980 ) was used with some modifications.

10 g of red ginseng extract obtained in the above example, its soluble extract, its fraction or flavonoid compound isolated therefrom, 4.0 l of microsome solution obtained in Experimental Example 2-1, activity assay buffer (0.5 M KH ₂ PO ₄ , 10 mM DTT, pH 7.4; Sigma) 20.0 μl, fat free bovine serum albumin (BSA; bovine serum albumin, stock solution 40 mg / ml; Sigma) 15.0 μl, cholesterol (stock solution 20 mg / ml 2.0 µl of Sigma) and 41.0 µl of distilled water were added and preliminarily reacted at 37 ° C for 15 minutes. 8 µl of [1- ¹⁴ C] oleoyl-coei (0.05 µCi, final concentration: 10 µM) was added to the reaction mixture and reacted again at 37 ° C. for 30 minutes, followed by an isopropanol: heptane mixture (4: 1 (v / v)) 1 ml was added to stop the reaction. To this was added 600 μl of heptane and 200 μl of 0.1 M KH ₂ PO ₄ (pH 7.4), the mixture was vigorously mixed with a vortexer and centrifuged at 300 rpm for 5 minutes. 100 μl of the supernatant obtained by centrifugation was placed in a scintillation bottle, and 4 ml of scintillation liquid (Lipoluma) was added thereto. The radiation dose of this mixture was measured with a 1450 Microbeta liquid scintillation counter (Wallacoy).

ACAT activity was calculated from the synthesized cholesteryl oleate picolol (picomol / min / mg protein) per mg of protein for 1 minute by calculating the radiation dose per hour from the measured radiation dose. The results are shown in Table 3 .

 Extract or compound Inhibition rate hACAT-1 (25 μg / ml) hACAT-2 (25 μg / ml) Ethanol extract 26.9% inhibition at 100 20.5% inhibition at 100 μg / ml Hydrothermal extract 18.6% inhibition at 100 / 14.8% inhibition at 100 μg / ml n-hexane soluble extract 23.5% inhibition at 100 11.5% inhibition at 100 μg / ml Chloroform Soluble Extract 63.2% inhibition at 100 49.5% inhibition at 100 μg / ml Ethyl acetate soluble extract 47.5% inhibition at 100 45.0% inhibition at 100 μg / ml 5,7,2 ', 4'-tetrahydroxy-8-rabbendulylflavanone 49.9 32.9 5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone 28.1 32.7 5,7,4'-trihydroxy-8-lavendulyl-2'-methoxyflavanone 59.4 40.0 5,7,2'-trihydroxy-8-lavendulylflavanone 72.6 52.2 5,7,2'-trihydroxy-8- (5-hydroxy-2-isopropenyl-5-methylhexyl) -flavanone 35.8 32.8 5,2'-dimethoxy-7,4'-dihydroxy-8-lavendulylflavanone 30.1 19.7

As shown in Table 3 , it was found that the ginseng extract, the soluble extract thereof, the fraction thereof or the flavonoid compound separated therefrom contained in the composition according to the present invention has excellent inhibitory ability against ACAT activity. Chloroform soluble extract and ethyl acetate soluble extract showed more than 40% inhibition at 100 ㎍ / ml on the activity of human ACAT-1 and human ACAT-2, especially 5,7,2'- which is a flavonoid compound. Trihydroxy-8-rabbendulyl flavanone inhibited the activity of human ACAT-1 by 70% or more, and inhibited the activity of human ACAT-2 by 50% or more.

<Experimental Example 3> Acute oral toxicity test for ICR mouse

In order to determine the acute toxicity of the ginseng extract, soluble extract thereof, fraction thereof or flavonoid compound isolated from the composition according to the present invention, the following experiment was carried out. Twelve female and 12 males (three males and three females each) of ICR mouse specific pathogens free at 4 weeks of age were housed in an animal room at a temperature of 22 ± 3 ° C, 55 ± 10% humidity and illumination of 12L / 12D. Breeding in. Mice were allowed to acclimate for about a week before being used in the experiment. Feed for experimental animals (Jeil Jedang Co., Ltd., mice and rats) and drinking water were supplied after sterilization and freely ingested.

The ginseng extract prepared in Example, its soluble extract, its fraction, or flavonoid compound isolated therefrom was prepared at a concentration of 50 mg / ml using 0.5% Tween 80 as a solvent, and then 0.04 ml (100 g of mouse weight) (100 g). mg / kg), 0.2 ml (500 mg / kg), 0.4 ml (1,000 mg / kg) orally. Samples were administered orally once and observed for side effects or lethality for 7 days after administration. That is, on the day of administration, changes in general symptoms and the presence or absence of dead animals were observed at least one hour, four hours, eight hours, twelve hours after the administration, and one or more mornings and afternoons every day from the day after the administration. In addition, on the 7th day of administration, animals were killed and dissected, and the internal organs were visually examined. Changes in body weight were measured at daily intervals from the day of administration to observe the weight loss phenomenon of the animal by the extract of Ginseng, its soluble extract, its fractions or the compounds isolated therefrom.

As a result, all mice treated with test substance showed no clinical symptoms and no dead mice, and no toxicity change was observed in weight change, blood test, blood biochemistry test and autopsy findings. Therefore, the ginseng extract according to the present invention, its soluble extract, its fraction and the compound isolated therefrom did not show toxicity change up to 1000 mg / kg in all mice, and the minimum lethal dose (LD ₅₀ ) of at least 1,000 mg / It was determined to be a safe substance that is more than kg.

Hereinafter, the formulation example for the composition of this invention is illustrated.

Preparation Example 1 Preparation of Pharmaceutical Formulation

1-1. Powder  Produce

2 g of one kind of flavonoid compound

1 g lactose

The above ingredients were mixed and filled in airtight cloth to prepare a powder. When using ginseng extract, soluble extract thereof or fractions thereof instead of the flavonoid compound, the extract or fraction is used to remove the solvent completely and diluted in purified water.

1-2. Manufacture of tablets

100 mg of one kind of flavonoid compound

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets. When using ginseng extract, soluble extract thereof or fractions thereof instead of the flavonoid compound, the extract or fraction is used to remove the solvent completely and diluted in purified water.

1-3. Preparation of Capsules

100 mg of one kind of flavonoid compound

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule. When using ginseng extract, soluble extract thereof or fractions thereof instead of the flavonoid compound, the extract or fraction is used to remove the solvent completely and diluted in purified water.

1-4. Preparation of Injection

10 ㎍ / mL 1 kind of flavonoid compound

Dilute hydrochloric acid BP until pH 3.5

Injectable sodium chloride BP up to 1 ml

The flavonoid compound was dissolved in an appropriate volume of sodium chloride BP for injection, the pH of the resulting solution was adjusted to pH 3.5 with dilute hydrochloric acid BP, and the volume was adjusted with sodium chloride BP for injection and mixed well. The solution was filled into a 5 ml Type I ampoule made of clear glass, dissolved under glass to be sealed under an upper grid of air, and sterilized by autoclaving at 120 ° C. for at least 15 minutes to prepare an injection solution. When using ginseng extract, soluble extract thereof or fractions thereof instead of the flavonoid compound, the extract or fraction is used to remove the solvent completely and diluted in purified water.

Preparation Example 2 Preparation of Food

Food containing a ginseng extract, a soluble extract thereof, a fraction thereof or a flavonoid compound separated therefrom was prepared as follows. At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

2-1. Preparation of Cooking Seasonings

Red ginseng extract, soluble extract thereof, fractions thereof, or flavonoid-based compounds isolated from 0.2 to 10% by weight of a health promotion cooking seasoning was prepared. At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

2-2. tomato ketchup  And preparation of sauces

Red ginseng extract, soluble extract thereof, fractions thereof, or 0.2 to 1.0 wt% of flavonoid compounds isolated therefrom was added to tomato ketchup or sauce to prepare tomato ketchup or sauce for health promotion. At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

2-3. Manufacture of Flour Food

0.1 to 5.0% by weight of red ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds isolated therefrom are added to flour, and bread, cake, cookies, crackers and noodles are prepared using the mixture to prepare foods for health promotion. Prepared. At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

2-4. soup  And juicy ( gravies Manufacturing

Red ginseng extract, soluble extract thereof, fractions thereof, or 0.1 to 1.0% by weight of flavonoid compounds isolated therefrom were added to soups and broth to prepare meat products for health promotion, soups and broths of noodles. The ginseng extract is used to remove the solvent completely and diluted in purified water.

2-5. ground Ground beef  Produce

Ginseng extract, soluble extract thereof, fractions thereof, or 10% by weight of flavonoid compounds isolated therefrom was added to ground beef to prepare ground beef. At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

2-6. dairy product( dairy products Manufacturing

0.1 to 1.0 wt% of red ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds separated therefrom were added to milk, and various dairy products such as butter and ice cream were prepared using the milk. At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

2-7. Linear  Produce

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh. Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh. The extract of red ginseng, its soluble extract, its fractions or flavonoid compounds isolated therefrom were concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by spraying and drying with a hot air dryer was ground to a particle size of 60 mesh to obtain a dry powder.

Cereals, seeds, and ginseng extracts, soluble extracts thereof, fractions thereof, or dry powders of flavonoid compounds prepared in the above ratio were prepared.

Cereals (30 wt% brown rice, 15 wt% brisk, 20 wt% barley)

Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame)

Dry powder of ginseng extract or flavonoid compound (1% by weight)

Ganoderma lucidum (0.5 wt%)

Foxglove (0.5 wt%)

At this time, the ginseng extract, its soluble extract or fractions thereof is used after diluting the solvent completely, diluted in purified water.

Preparation Example 3 Preparation of Beverage

3-1. Preparation of Carbonated Drinks

5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed, and 79-94% purified water is mixed to make syrup, and the syrup is 85-98 Sterilized at ℃ for 20 ~ 180 seconds, mixed with cooling water at a ratio of 1: 4, and then injected with 0.5 to 0.82% of carbon dioxide gas to produce a carbonated beverage containing red ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds separated therefrom. Was prepared. The ginseng extract, its soluble extract, or its fraction is used to remove the solvent completely and diluted in purified water.

3-2. Health drink  Produce

Substances such as liquid fructose (0.5%), oligosaccharides (2%), sugars (2%), salts (0.5%), water (75%), and ginseng extracts, soluble extracts thereof, fractions thereof or flavonoids separated therefrom The compound was homogeneously mixed and sterilized immediately, and then packaged in a small packaging container such as a glass bottle or a plastic bottle to prepare a health beverage. The ginseng extract, its soluble extract, or its fraction is used to remove the solvent completely and diluted in purified water.

3-3. Vegetable juice  Produce

0.5 g of red ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds isolated therefrom were added to 1000 ml of tomato or carrot juice to prepare vegetable juice for health promotion. The ginseng extract, its soluble extract, or its fraction is used to remove the solvent completely and diluted in purified water.

3-4. Fruit juice  Produce

0.1 g of red ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds isolated therefrom were added to 1000 ml of apple or grape juice to prepare fruit juice for health promotion. The ginseng extract, its soluble extract, or its fraction is used to remove the solvent completely and diluted in purified water.

Claims (13)

A composition for the prevention and treatment of cardiac circulatory disorders, comprising a ginseng extract, a soluble extract thereof, a fraction thereof or a flavonoid compound isolated therefrom as an active ingredient. The method of claim 1, wherein the extract of ginseng is a composition for preventing and treating cardiovascular diseases, characterized in that the extract is prepared by extracting the ginseng with water, C ₁ ~ C ₄ alcohol, or a mixed solvent thereof. According to claim 2, wherein the alcohol is methanol, ethanol or butanol composition for the prevention and treatment of cardiovascular diseases, characterized in that. According to claim 1, The ginseng extract is a composition for the prevention and treatment of cardiovascular diseases, characterized in that produced by extracting the ginseng with hot water. The method of claim 1, wherein the soluble extract is concentrated to remove the ginseng extract to completely remove the solvent, suspended in water, alcohol of C ₁ ~ C ₄ , or a mixed solvent thereof, sequentially with n-hexane, chloroform and ethyl acetate N-hexane soluble extract, chloroform soluble extract or ethyl acetate soluble extract obtained by fractionation as a composition for the prevention and treatment of cardiovascular diseases. 6. The composition for preventing and treating cardiovascular diseases according to claim 5, wherein the suspension solvent is a mixed solvent of water and methanol. According to claim 1, wherein the fraction is 100% chloroform, n-hexane / acetone (100: 1 to 1: 5, volume ratio) mixed solvent and chloroform / acetone (150: 1 to 1: 4, volume ratio relative to the soluble extract ) A composition for the prevention and treatment of cardiovascular diseases, characterized in that obtained by performing a silica gel chromatography using a mixed solvent as a mobile phase. 8. The composition for preventing and treating cardiovascular diseases according to claim 7, wherein the soluble extract is a chloroform soluble extract. The method of claim 1, wherein the flavonoid compound 1) 5,7,2 ', 4'-tetrahydroxy-8-rabbendulylflavanone; 2) 5-methoxy-7,2 ', 4'-trihydroxy-8-rabbendulylflavanone; 3) 5,7,4'-trihydroxy-8-rabbendulyl-2'-methoxyflavanone; 4) 5,7,2'-trihydroxy-8-rabbendulylflavanone; 5) 5,7,2'-trihydroxy-8- (5-hydroxy-2-isopropenyl-5-methylhexyl) -flavanone; And 6) 5,2'-dimethoxy-7,4'-dihydroxy-8-rabbendulyl flavanone The composition for the prevention and treatment of cardiovascular diseases, characterized in that at least one member selected from the group consisting of. The method of claim 1, wherein the cardiovascular disease is hyperlipidemia, coronary heart disease, arteriosclerosis or myocardial infarction, characterized in that the composition for the prevention and treatment of cardiovascular diseases. According to claim 1, The ginseng extract, soluble extract thereof, fractions thereof or flavonoid compounds isolated from the composition for the prevention and treatment of cardiovascular diseases, characterized in that it has an antioxidant activity. According to claim 1, The ginseng extract, soluble extracts thereof, fractions or flavonoid-based compounds isolated therefrom is acyl coay: cholesterol acyl transferase inhibitory composition, characterized in that for preventing and treating cardiovascular diseases . Claim 1, ginseng extract, soluble extract thereof, fractions thereof, or a flavonoid-based compound separated therefrom as an active ingredient food composition for improving heart circulation disease.