KR20110133931A - Pharmaceutical composition comprising an extract of chokeberry for preventing and treating arteriosclerosis and hypertension - Google Patents

Abstract

PURPOSE: A pharmaceutical composition and food composition containing C3G complex(chokeberry biologically active fraction) is provided to ensure safety and to minimize side effect. CONSTITUTION: A pharmaceutical composition for preventing and treating arteriosclerosis contains 15% or more of C3G complex(cyanidin-3-glucoside complex) as an active ingredient. A pharmaceutical composition for preventing and treating essential hypertension contains C3G complex(chokeberry biologically active fraction) as an active ingredient. The C3G complex is administered in 1-200 mg/kg. A food composition for preventing and treating arteriosclerosis contains the C3G complex as an active ingredient.

Description

Pharmaceutical composition comprising an extract of chokeberry for preventing and treating arteriosclerosis and hypertension containing chokeberry bioactive fraction C3K complex as an active ingredient

The present invention provides a pharmaceutical composition for preventing and treating arteriosclerosis and hypertension, which contains C3G complex (Cyanidin-3-glucoside complex, chokeberry bioactive fraction) as an active ingredient. More specifically, the present invention can inhibit the onset of atherosclerosis produced by a high-fat, high-lipid diet in the aortic vessel, and contain a C3G complex (chokeberry bioactive fraction) capable of lowering essential hypertension as an active ingredient. It relates to a pharmaceutical composition and food composition for preventing and treating atherosclerosis and hypertension.

In recent years, rich and diverse diets and lifestyle changes have made nutrition intake prone to imbalance and mechanized modern life has resulted in lack of exercise. As a result, the prevalence of chronic diseases such as hypertension and hypertension is increasing, and the form of the disease is changing to an advanced country type. 27.7% of the total deaths of men and women after Korea in their 40s (National Statistical Office 2004) are dying from circulatory diseases, and the prevalence of circulatory diseases increases gradually to 324 (3.2%) per 10,000 (National Health Insurance Service, 2000). .

Coronary artery disease (coronary artery disease) refers to a disease in which myocardial ischemia may occur due to impaired blood supply to the myocardium due to coronary artery stenosis caused by atherosclerosis. Atherosclerosis is characterized by the deposition of lipids into the arterial wall by damage to vascular endothelial cells and the release of endothelial cells, including macrophages, T-lymphocytes and smooth muscle cells and cytokines, growth factors, etc. React with each other and progress to atherosclerosis. As such, immune cells are almost involved in the progression of atherosclerosis, and because of the similarities between atherosclerosis and inflammatory diseases such as rheumatoid arthritis and pulmonary fibrosis, the interpretation of atherosclerosis is dominant in recent years.

One of the major risk factors for atherosclerotic cardiovascular disease is hyperlipidemia. Inhibitors of 3-hydroxyl-3-methylglutaryl coenzyme A (HMG-CoA) reductase in large-scale clinical trials in patients with coronary heart disease. The statin regimen did a very good job. Statins are structurally similar to HMG-CoA, the precursor of cholesterol, and competitively inhibit HMG-CoA reductase activity. However, the lipid-lowering effect alone is insufficient to explain the therapeutic and prophylactic effects of statins on atherosclerotic cardiovascular disease. Many clinical trials and basic scientific studies have shown that the anti-inflammatory properties of statins inflammatory properties are revealed. In other words, the anti-inflammatory effect is emerging as the most important factor in treating atherosclerosis.

On the other hand, the heart pumps constantly to circulate blood in our body. The pressure on the walls of blood vessels is called blood pressure. In general, the blood pressure contains pressure when the heart contracts and the blood expands to contain blood. There are two pressures. In general, blood pressure varies according to an individual's physique, age, man, woman, stress, and season. Hypertension is one of five Korean adults with hypertension. Hypertension is a frightening adult disease that progresses without symptoms and suddenly leads to arteriosclerosis, which again causes fatal complications such as stroke and myocardial infarction. Heart or brain. If you don't have any symptoms before complications occur in various organs such as the kidneys, you can get worse if you miss the initial treatment.

Normal blood pressure usually refers to systolic blood pressure of 140 or less and diastolic blood pressure of 90 or less, whereas hypertension refers to systolic blood pressure of 160 or more and diastolic blood pressure of 95 or more. High blood pressure should be checked for secondary hypertension, the presence of other risk factors for atherosclerosis, and the extent of damage to major organs.

Hypertension is divided into two types depending on the cause. Essential hypertension is unclear in cause and tends to be hereditary and usually occurs at the age of 20s to 50s. About 80% of hypertensive patients in Korea belong to this group, and the close relationship between hypertension and genetic factors. The other is secondary hypertension, which is preceded by the causative disease and one of the symptoms of the disease is hypertension. It originates from the environment and lifestyle, causing smoking, drinking, stress, obesity, and excessive salt intake. Salt and obesity in particular are the main causes of high blood pressure.

Hypertension is mainly explained by the physiological mechanism of the renin-angiotensin system (RAS), which is mainly distributed in the kidneys and plays a very important role in the creation and maintenance of hypertension, such as regulating renal function and blood pressure. In addition, it is recently known to be distributed in various tissues such as heart, liver, lungs, etc., the importance of which is emerging. The most important enzyme in the regulation of RAS is angiotensin converting enzyme (ACE), which converts angiotensin to bioactive angiotensin Ⅱ (ang Ⅱ).

Ang II is a substance directly involved in blood pressure elevation by increasing blood vessel contraction and aldosterone secretion by binding to angiotensin II-1 type receptor (type 1 (AT1))-receptor, and superoxide anion and hydrogen peroxide. Increasing the production of reactive oxygen species (ROS) and reactive nitrogen species (RNOS) such as peroxide, hydroxyl radicals, and peroxynitrites, resulting in functional gas phase and LDL oxidation of endothelial cells. Promotes the inflammatory response of blood vessels.

Ang II also causes oxidative stress in endothelial cells through the action of NADH / NADPH oxidase. Stimulation of this enzyme in Spontaneously hypertensive rats (SHR) induces the production of vascular superoxide ions. It reduces the bioavailability of NO by causing dysfunction of endothelial cells such as hypertrophy and death. In addition, ACE also acts on the kallikrein / kinin system to decompose the C-terminus of bradykinin, a vascular relaxant, to increase blood pressure by inhibiting vascular relaxation. Antioxidant action in these RAS and ROS-related mechanisms interferes with the action of Ang II and AT1-receptors, increases the activity of antioxidant enzymes, reduces cellular damage, increases NO production, and reduces the risk of hypertension through vasodilation. Let's go. In other words, it is believed that a substance with high antioxidant activity may have an effect of preventing and treating arteriosclerosis and hypertension.

Since ACE is distributed in various tissues such as lungs, heart and kidneys and plasma, ACE inhibitors have been developed to lower blood pressure by inhibiting Ang II production and degradation of bradykinin. Enalapril, captopril, zofenopril, lisinopril and ternocapril have been developed and are now commercially available, including high blood pressure, congestive heart failure, It is widely used as a treatment for myocardial infarction, diabetic nephritis and kidney failure. However, these drugs have many side effects such as taste disorders, white blood cell reduction, angioedema, liver dysfunction, and dry cough, and many studies are being actively conducted to develop ACE inhibitors from natural substances.

In addition, the antihypertensive drugs currently widely used in clinical practice have many side effects. Among them, diuretic-based hypertension drugs cause hypocalcemia, hyperuremic acid, hyperlipidemia, sexual dysfunction, and sympathetic neuropathic blood pressure lowering agents appear sexual dysfunction, thirst, depressive bradycardia. Vasodilating hypotensive drugs include tachycardia, angina pectoris, helplessness, and flushing. Recently developed Ca ²⁺ antagonists have almost established efficacy, but have side effects such as inhibition of cardiac function due to myocardial contraction, and insulin secretion disorder. It is known to cause. Many high blood pressure medications suffer from the serious side effects of using these side effects while still being used. Therefore, the necessity of developing natural medicines or new drugs to improve the harm caused by these side effects is emerging.

Currently, many studies are being conducted around the world to use drugs extracted from natural products for the prevention or treatment of diseases. Among them, there are many reports that berry extract has antioxidant and antibacterial effects. In particular, chokeberries grown in large quantities in Poland have a high content of antioxidants such as anthocyanins, catechins, and chloric acid, which are phenolic compounds that exhibit antioxidant effects. It has been reported that there is a positive effect on the inhibition of endothelial cell apoptosis, growth of vascular endothelial progenitor cells and anti-inflammatory effect.

However, C3G complexes (chokeberry bioactive fractions) can inhibit the development of atherosclerosis produced by a high fat, high-lipid diet and lower essential hypertension, which can be used for the treatment and prevention of atherosclerosis and hypertension. No disclosure or teaching has been made.

The inventors of the present invention can suppress the onset of atherosclerosis produced by a high-fat, high-lipid diet in the aortic vessel of the C3G complex (chokeberry bioactive fraction), a substance extracted and purified from chokeberry fruit at high purity, and lowers essential hypertension. It was confirmed that the present invention was completed.

Accordingly, an object of the present invention is to provide a pharmaceutical composition and a food composition for preventing and treating arteriosclerosis and hypertension, which contain a C3G complex (chokeberry bioactive fraction) as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention and treatment of atherosclerosis produced by a high fat high lipid diet containing a C3G complex (chokeberry bioactive fraction) as an active ingredient.

To determine whether the atherosclerotic model animal, Homozygotic apolipoprotein E knock out (ApoE) mice, can suppress the development of atherosclerosis produced by high fat high fat diet in the aortic vessels, ApoE mice were treated with high fat high cholesterol diet. Observation of the progression of the lesions by oral administration of C3G conjugates during the induction process effectively inhibited the infiltration of inflammatory cells into the atherosclerotic lesions of the inflammatory cells with only two administrations per week (FIGS. 3A and 3A). 3b). In addition, the suppression of LPS (Lipopolysaccharides) -induced MCP-1 (Monocyte chemoattractant protein 1) in macrophages is a clear explanation of the invasion inhibition mechanism in animals. In addition, the results showed that the effective capturing ability of the radical can be more effective than vitamin C.

In addition, the expression of SREBP-1 (Sterol Regulatory Element Binding Protein-1) and LPL (lipoprotein lipase), which are lipid metabolism-related genes of macrophages, was effectively inhibited by C3G conjugates, and the C3G complex (chokeberry bioactive fraction). It was also confirmed that the plasma TG concentration of the treated mice group was significantly lower (FIG. 5). These results suggest that C3G is a potential anti-arteriosclerosis drug.

According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing and treating essential hypertension, which contains a C3G complex (chokeberry bioactive fraction) as an active ingredient.

To determine whether SHR, a naturally occurring hypertension mouse, can be used to reduce the high hypertension induced by angiotensin II, high blood pressure after administration of C3G in high blood pressure rats at high risk of cardiovascular disease. Blood pressure was significantly lower than that of the control group (see FIG. 7), and left ventricular hypertrophy was corrected (see Table 1). Blood TG levels were partially decreased in the C3G complex (chokeberry bioactive fraction) treated group than in the control group, and glucose was significantly decreased in the C3G conjugated group. Fibrosis and thickening in the heart and kidneys due to the amount of collagen deposition were reduced in the C3G group. These results were significantly lowered in blood pressure compared to the amlodipine group, which is a commonly used calcium antagonist for the treatment of hypertension.

The chokeberry is reddish purple and contains the most anthocyanins and polyphenols among the existing berry fruits. The scientific name of chokeberry is known as Aronia melanocarpa or Pyrus melanocarpa . It is a perennial shrub rosaceae, and its anthocyanin content is known to be 5 times higher than blueberries, 10 times more than cranberries, and 20 times higher than bokbunja. Such chokeberry is known to have anti-cancer effects, anti-inflammatory effects, and spermatosis (reduction) through various clinical trials.

In the present invention, the C3G complex (chokeberry bioactive fraction) has a cyanine content of 15% or more, and is composed of cyanine, anthocyanidin, phenolic acid, and the like.

In the pharmaceutical composition according to the present invention, the C3G complex (chokeberry bioactive fraction) may preferably be contained in an amount of 0.0001 to 30% by weight, more preferably 0.001 to 10% by weight, based on the total weight of the composition. It may be contained in an amount of. If the content is less than 0.0001% by weight, it is difficult to obtain the desired effect in the present invention, and if it exceeds 30% by weight, the content is preferable as described above because it causes a problem in the stability of the formulation.

The composition of the present invention can be used in the form of a pharmaceutically acceptable salt, and acid salts formed by pharmaceutically acceptable free acid are useful as salts. The C3G complex can form pharmaceutically acceptable acid addition salts according to methods conventional in the art. Free acid and inorganic acid may be used as the free acid, and hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, and fumaric acid may be used as the organic acid. (fumaric acid), formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galluxuric acid, embonic acid, glutamic acid or aspartic acid Can be used.

In the composition of the present invention, the C3G complex may be extracted using water, an organic solvent or a mixed solvent thereof, which is a well-known extraction solvent for conventional natural product extraction, but preferably an active substance present in a large amount in the shell after compression extraction. It is characterized in that extracted with ethanol. According to the above method, there is an advantage that the damage of the pure active ingredient is very small and the yield is high.

The compositions of the present invention may be formulated in the form of powders, granules, tablets, capsules, injections, creams, gels, patches, sprays, drinks or ointments by conventional methods known in the art. Moreover, in order to make storage and handling easy, you may add the carrier used for normal formulation, such as dextrin and cyclodextrin, and other arbitrary preparations.

The pharmaceutical composition containing the C3G complex of the present invention as an active ingredient may further include appropriate carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

In addition, carriers, excipients and diluents that may be included in the composition containing the C3G complex as an active ingredient include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, Alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. . When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the C3G complex, for example, starch, calcium carbonate, sucrose. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the C3G complex of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and can be appropriately selected by those skilled in the art. However, for the desired effect, the C3G complex of the present invention is preferably administered at 0.0001 to 1000 mg / kg on a daily basis, 1 to 200 mg / kg to be more effective. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

C3G complexes of the invention can be administered by a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

According to another aspect of the present invention, the present invention provides a food composition for preventing and improving atherosclerosis containing a C3G complex (chokeberry bioactive fraction) as an active ingredient.

According to another aspect of the present invention, the present invention provides a food composition for preventing and improving hypertension containing C3G complex (chokeberry bioactive fraction) as an active ingredient.

In the food composition according to the present invention, the C3G complex (chokeberry bioactive fraction) may be contained in an amount of 0.0001-30% by weight based on the total weight of the composition, and when it is less than 0.0001% by weight, it is difficult to obtain the desired effect in the present invention. It is difficult and, if it exceeds 30% by weight, such a content is preferable because of a problem in the stability of the formulation.

In the composition of the present invention, the C3G complex may be extracted using water, an organic solvent, or a mixed solvent thereof, which is a well-known extraction solvent for conventional natural product extraction, but is preferably a compressed and ethanol extract.

In the present specification, the term "food" means a natural product or processed product containing one or more nutrients, and preferably means a state in which it can be directly eaten through some processing process. Includes food, food additives, nutraceuticals and beverages.

The food of the present invention may further comprise a food acceptable food supplement, and may be formulated in the form of pills, powders, granules, acupuncture, tablets, capsules or beverages.

In addition, the composition of the present invention can be used as a food supplement of health supplements for the prevention of related diseases. Foods to which the C3G complex of the present invention may be added include various foods, for example, beverages, gums, teas, vitamin complexes, dietary supplements, and the like, pills, powders, granules, tablets, capsules, or beverages. Can be used in the form of phosphorus.

In this case, the amount of the C3G complex in the food or beverage can be generally added to 0.0001 to 30% by weight of the total food weight in the case of the food composition of the present invention, 0.0001 to 20% by weight based on 100ml for the health beverage composition Can be added.

Food additives other than the C3G complex as defined in the present invention include food additives customary in the art, such as flavors, flavors, colorants, fillers, stabilizers and the like, and are exemplified below.

In the case of the beverage composition, there are no particular limitations on the liquid component other than the C3G complex as an essential component in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional components, as in general beverages. Examples of the natural carbohydrates include, for example, monosaccharides such as glucose and fructose, for example, disaccharides such as maltose and sucrose, for example, conventional sugars such as polysaccharides such as dextrin and cyclodextrin, and xylitol. Sugar alcohols such as sorbitol and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.001 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

As described above, the chokeberry bioactive fraction according to the present invention, the C3G aggregate, can suppress the onset of arteriosclerosis produced by a high fat high lipid diet in the aortic vessel, and can lower the intrinsic hypertension, thereby reducing arteriosclerosis and It is expected to be of high value as a natural resource derived from natural products that can be used for the purpose of preventing and treating essential hypertension.

1 is a diagram schematically showing an experimental protocol for investigating the effect on atherosclerosis of the C3G complex according to the present invention.
Figure 2a is a photograph showing the effect of C3G conjugates according to the present invention on lipid deposition in the aorta (Aorta).
2B is a graph showing the results of statistical processing of lipid deposition in the aorta (Aorta).
3A is a photograph showing lipid deposition in the aortic arch.
3B is a graph showing the results of statistical treatment of lipid deposition in the aortic arch.
Figure 4a is a photograph showing the invasion of aortic arch macrophages.
Figure 4b is a graph showing the results of statistical processing of invasion of aortic arch macrophages.
5 is a diagram showing that the plasma TG concentration of the C3G complex (chokeberry bioactive fraction) treated mice group of the present invention is significantly lowered. (TG: Triglyceride, TC: Total Cholesterol, LDL: Low Density Protein Cholesterol, HDL: High Density Protein Cholesterol)
Figure 6 schematically shows an experimental protocol for examining the effect of the C3G complex according to the present invention on hypertension.
7 is a diagram showing the effect of the C3G complex according to the present invention on the systolic blood pressure of the hypertensive rats (SHR). (* p <0.05 Berry and amlodipine-treated vs. vehicle-treated SHR)

Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1. Preparation of C3G Complex

1) M. chokeberry (Marek chokeberry) fruit was purchased and stored at -15 degrees frozen.

2) Frozen M. chokeberry fruit was thawed at -2 degrees.

3) After compression extraction (190 ~ 290 r / min, 0.5 ℃), M. chokeberry extract was transferred to the buffer container and the residue was transferred to the extractor. In the extractor, the residue was periodically extracted with 50% ± 2% ethanol. The process took place at 15-25 ° C. The obtained ethanol extract was collected in a buffer container and transferred to a thin film evaporator for concentration and ethanol evaporation.

4) Decantation (Choking pressure 2 ~ 2.5 bar, 300 ~ 700dm3 / h) precipitate was removed. The precipitate of M. chokeberry extract was adjusted to 7% or less.

5) The decantation extract in Alfa -Lawal disk type separator was clarified.

6) The clarified extract was applied to a separation column of SPE (Solid Phase Extraction) method. The M. chokeberry extract was passed at 500 L / h and the column was washed with demineralized ionized water.

7) The polyphenol attached to the column was desorbed with 50% ethanol and the eluted ethanol eluate was collected in a buffer container.

8) The ethanol eluate was distilled and concentrated by thin film evaporator (-85 bar, 52.5 ° C.)

9) The concentrate was dried in a SRN pulverizing dryer. (Water content of powder is less than 10%)

10) Standardized powder was packaged in polypropylene.

Example 2. Composition and Content

1 kg of the test sample in the powder made in Example 1 was used for the experiment. The C3G complex used in the experiment was composed of cyanine, proanthocyanidin, and phenolic acid, which is a kind of anthocyanin, and the content of cyanine as a reference indicator was 15% or more.

Example 3 Antiarteriosclerosis Effect of C3G Suture

In the present invention, C3G complex (chokeberry bioactive fraction), that is, C3G complex containing high anthocyanin, which is a representative substance of phenolic compound known to have anti-inflammatory effect, has an effect on atherosclerotic cardiovascular disease In order to confirm the effect, the arteriosclerosis animal model was used.

1) Animal test preparation (see experimental protocol of FIG. 1)

Ten mice were assigned to each group as defined according to the defined experimental protocol.

Group 1: Normal diet group

Group 2: High Fat High Cholesterol Diet Group (Contains 0.15% Cholesterol)

Group 3: High fat, high cholesterol diet + C3G diet 20 mg / kg joint diet group

Group 4: High fat, high cholesterol diet + C3G diet 100 mg / kg joint diet group

Group 5: high fat, high cholesterol diet + C3G diet 200 mg / kg joint diet group

Each group was orally administered a fixed dose of C3G conjugate twice a week for 8 weeks.

2) Inhibition of Lipid Deposition in Aorta and Aortic Arch by C3G Synthesis

Eight weeks later, mice were dissected to investigate whether administration of C3G conjugates affected the apoptosis produced when a high-fat, high-cholesterol (HFHC) diet was administered to ApoE mice used as an arteriosclerosis experimental animal model. It was. The aorta of the mouse was isolated, quenched and frozen in an OCT compound (OCT coumpound), and sections of the tissues were prepared and stained with oil red O (oil red O). Red stained lipids were significantly higher in mice fed the HFHC diet compared to the control mice, at which time the C3G co-administrated group showed a decrease, but the results were not significant (Fig. 2b, p). = 0.1, 0.17).

In addition, oil red O staining of the aortic sinus sections of these mice also significantly increased lipid deposits on the arterial wall of mice fed the HFHC diet, and the group treated with C3G mixtures. It was shown that the degree of lipid deposition in the dose-dependently decreased concentration of C3G conjugate (FIG. 3A).

As a result of statistically processing the results obtained from 9 mice in each group, the degree of deposition was significantly reduced in the group administered with 200 mg / kg of C3G conjugate (FIG. 3B, p = 0.08).

3) Inhibition of Macrophage Invasion in Aortic Arch by C3G Synthesis

Recently, since the view that arteriosclerosis is an inflammatory disease is generally accepted, H & E staining of the fixed aortic sinus cross section showed that the infiltration of inflammatory cells was reduced in the group co-administered with C3G.

Thus, immunostaining was performed with MOMA-2 antibodies to determine the infiltration of macrophages, which are reported to be the most important of inflammatory cells. As shown in Figure 4, while severe macrophage infiltration into the artery arch of the HFHC-fed rats was observed, the subjects in combination with the C3G conjugate can see a marked decrease in macrophages infiltrated into the atherosclerotic lesions. In particular, in the group treated with C3G aggregates, the infiltration was reduced in a concentration-dependent manner (FIG. 4A), and the statistical results were also significantly reduced (FIG. 4B). In particular, in the case of the group administered with the concentration of C3G 200 mg / kg it was confirmed that the marked decrease in macrophage invasion (Fig. 4b, p <0.01).

Administration of only two C3G supplements per week for eight weeks altered the appearance of macrophages most important for the development of atherosclerosis and effectively inhibited the infiltration of inflammatory cells into atherosclerotic lesions in the atherosclerosis. It can be said that the results show the potential as an anti-arteriosclerosis treatment.

Example 4. Effect on the prevention and treatment of hypertension of C3G conjugate

In the present invention, SHR (Spontaneously Hypertensive Rats), a model for spontaneous hypertension disease with concomitant blood pressure rise and development of rats, was used. This model is a disease research model that is commonly used to study essential hypertension in humans after 9 weeks of elevated systolic blood pressure of more than 180 mmHg. In the present invention, the experiment was initiated with 7-week-old SHR to confirm the development and accompanying blood pressure increase, and to examine the efficacy of the prevention and treatment of hypertension of C3G co-forms through animal experiments. The experimental group was a high blood pressure control group in which only drinking water was administered to SHR, the Sham group, the C3G group 100 mg / kg group, the C3G group 200 mg / kg group, and the calcium channel blocker commonly used as a general hypertension drug, amlodipine. ) Was divided into four experimental groups as a normal control group administered 10 mg / kg. C3G conjugate and amlodipine (amlodipine) were dissolved in drinking water by concentration and administered orally for a total of 8 weeks every day (see FIG. 6).

After 8 weeks, the blood pressure lowering effect of C3G was as much as the blood pressure (187.4 ± 22.7mmHg) of the SHR group who received 10mg / kg (am) of amlodipine, a typical drug for lowering blood pressure as a calcium channel blocker. Although not lowered, both concentrations were significantly different from the hypertensive controls. The systolic blood pressure of the SHR group treated with 100 mg / kg C3G was significantly reduced to 202.2 ± 10.2 mmHg, compared with 224.4 ± 12 mmHg in the control sham group. The group receiving 200 mg / kg C3G was significantly reduced to 202.0 ± 12.6 mmHg. In particular, the blood pressure lowering effect by the C3G aggregate was kept low after increasing the concentration of each from 100mg / kg to 200mg / kg from 30mg / kg to 100mg / kg (see Figure 7).

The tissue weights of the experimental groups sacrificed 8 weeks after the end of the experiment were measured. The weight-adjusted cardiac hypertrophy was 3.52 ± 0.1 mg in the sham hypertension control group, but decreased to 3.27 ± 0.09 at 100 mg / kg C3G and 3.30 ± 0.06 at 200 mg / kg C3G. In particular, the left ventricular hypertrophy was 2.85 ± 0.1 mg in the control group of sham hypertension, whereas the left ventricular hypertrophy was decreased in the group receiving the C3G combination of 2.68 ± 0.07 200 mg / kg in the 100 mg / kg C3G group and 2.68 ± 0.04 in the C3G group. Visible (Table 1).

In summary, after the administration of C3G conjugates in hypertensive rats at high risk of cardiovascular disease, blood pressure was significantly lowered compared to the hypertensive control group, and left ventricular hypertrophy was corrected. In conclusion, C3G is considered to be of high value as a natural plant resource that can be used for the purpose of preventing essential hypertension.

Formulation Example 1 Manufacturing Method of Syrup

A syrup containing the C3G complex of the present invention and a pharmaceutically acceptable salt thereof as an active ingredient of 2% (weight / volume) is prepared by the following method.

Acid addition salt, saccharin, and sugar of the C3G complex of the present invention were completely dissolved in 80 g of warm water, cooled, and then prepared by mixing with a solution consisting of glycerin, saccharin, spices, ethanol, sorbic acid and distilled water. Water was added to the mixture to a total volume of 100 ml. The addition salt can be replaced with other salts according to the examples.

The components of the syrup are as follows.

C3G complex, phosphate 2 g

Saccharin 0.8 g

25.4 g per

Glycerin 8.0 g

0.04 g of spices

Ethanol 4.0 g

0.4 g of sorbic acid

Distilled Water Determination

Preparation Example 1 Preparation of Health Functional Food

300 mg of C3G complex

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients may be mixed according to a general health functional food manufacturing method. Next, the granules may be prepared and used for preparing the health functional food composition according to a conventional method.

Although specific portions of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific techniques are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. It is therefore intended that the scope of the present invention be defined by the appended claims and their equivalents.

Claims (6)

A pharmaceutical composition for the prevention and treatment of atherosclerosis produced by a high fat and high lipid diet containing C3G complex (Cyanidin-3-glucoside complex, chokeberry bioactive fraction) as an active ingredient. [Claim 2] The pharmaceutical composition for preventing and treating atherosclerosis of claim 1, wherein the C3G complex (chokeberry bioactive fraction) has a cyanine content of 15% or more. Pharmaceutical composition for the prevention and treatment of essential hypertension containing C3G complex (chokeberry bioactive fraction) as an active ingredient. The pharmaceutical composition for preventing and treating essential hypertension of claim 3, wherein the C3G complex (chokeberry bioactive fraction) is administered at 1 to 200 mg / kg. Food composition for the prevention and improvement of atherosclerosis produced by a high fat high lipid diet containing C3G complex (chokeberry bioactive fraction) as an active ingredient. Food composition for preventing and improving essential hypertension containing C3G complex (chokeberry bioactive fraction) as an active ingredient.

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