WO2013133677A1

WO2013133677A1 - Composition containing reaction mixture of red-ginseng extract and persimmon vinegar for preventing or treating vascular diseases

Info

Publication number: WO2013133677A1
Application number: PCT/KR2013/001921
Authority: WO
Inventors: 박종대; 이경희; 정대영
Original assignee: 주식회사 천지양
Priority date: 2012-03-09
Filing date: 2013-03-11
Publication date: 2013-09-12
Also published as: KR101222845B1

Abstract

The present invention relates to a composition containing a reaction mixture of red-ginseng extract and persimmon vinegar for preventing or treating vascular diseases, the composition comprising augmented ginsenoside Rg₃ and arginine derivatives. The composition containing a reaction mixture of red-ginseng extract and persimmon vinegar may exhibit the superior effects in increasing NO generation, superior effects in inhibiting angiotensin-converting enzyme, and the like, and therefore can be useful as a composition for effectively preventing or inhibiting vascular diseases, including arteriosclerosis, hypertension, angina, myocardial infarction, ischemic heart disease, heart failure, complications caused after a transluminal coronary angioplasty, cerebral infarction, cerebral hemorrhage, strokes, etc.

Description

Composition for the prevention or treatment of vascular diseases containing the persimmon vinegar reaction mixture of red ginseng extract

The present invention relates to a composition for the prophylaxis or treatment of vascular diseases containing the persimmon vinegar reaction mixture of ginsenoside Rg ₃ and arginine derivative-enhanced red ginseng extract.

In modern society, vascular disease is a serious social problem. The vascular diseases include atherosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, heart failure, complications that occur after carotid angioplasty, cerebral infarction, cerebral hemorrhage, and stroke. According to the statistics of the causes of death published by the National Statistical Office in 2010, vascular diseases including hypertension, ischemic heart disease and cerebrovascular disease are the second leading causes of death in Korea, ranking second only to malignant tumors, and men are 55 years old. In women above 65, the mortality rate of vascular disease increases significantly. In particular, risk factors associated with atherosclerosis are age, sex, hypertension, hyperlipidemia, diabetes, smoking, lack of exercise and obesity. Patients with vascular disease receive constant medications to treat them, but they are not easy to cure in modern medicine. In addition, the longer the drug is administered, the greater the side effects.

Among these, hypertension is an increase in arterial blood pressure due to an increase in cardiac output or constriction of peripheral blood vessels. The low incidence of a single disease is caused by stroke, myocardial infarction, diabetes, and heart failure. It is enlarged and is also associated with the development of atherosclerosis. The pressure of blood flow through the blood vessels, ie arterial pressure, is called blood pressure. For some reason, systolic blood pressure is greater than 120 mmHg (maximum blood pressure) and diastolic blood pressure is greater than 80 mmHg (lowest blood pressure). Hypertension is divided into essential hypertension and secondary hypertension, which accounts for more than 90%. In the case of essential hypertension, there is no specific cure. This classification of high blood pressure not less than the activity of the sympathetic nervous system, the function of the cell membrane described above, water and a sodium (Na) excretion disorders of the _{_{kidney, PGE 2 (prostaglandin E 2)}} , PGG 12 (prostaglandin G 12), (atrial natriuretic peptide) ANP And hypotension of hypotensive agents such as endothelium dependent relaxing factor (EDRF), or high blood pressure control functions such as catecholamine and the renin-angiotensin system (RAS). have. Angiotensin converting enzyme (ACE), an important enzyme in the Lenin-Angiotensin system, one of the important mechanisms of blood pressure elevation, is an enzyme that converts angiotensin I and AngII into angiotensin II and AngII. It is known that the converted angiotensin II acts on the angiotensin II-type 1 receptor and constricts blood vessels and increases the release of aldosterone, thereby raising blood pressure (Am. J. Cardiol., 2003, 91 , 30-37). Meanwhile, among the endothelium-derived relaxing factors (EDRFs) in vascular endothelial cells, nitric oxide (NO) is known as the most important factor (Pro. Nat. Acad. Sci. USA, 1987, 84, 9265). ~ 9269). NO, which is released from vascular endothelial cells, has the effect of inhibiting platelet aggregation and adhesion in addition to vascular smooth muscle relaxation, thereby regulating blood circulation. NO suggests that NO is one of the most active researches in the medical community, suggesting that vascular endothelial cell damage and dysfunction are closely related to ischemic heart disease such as myocardial infarction and angina or adult diseases such as hypertension.

Meanwhile, angiotensin converting enzyme inhibitors such as enalapril, captopril, ramipril, and lisinopril have been developed and commercialized for the purpose of improving vascular diseases such as hypertension. The use of the drug has been reported side effects such as taste disorders, white blood cell reduction, angioedema, liver failure and dry cough (Elsevier, Amsterdam, 1984, 5, 198-199, 262-263, 298-300). In addition, it can be said that it is necessary to search for substances having a therapeutic effect on vascular diseases from natural substances having safety.

For the prevention and treatment of vascular diseases, many herbal medicines have been introduced in traditional textbooks such as folk or Dongbogam. Among them, red ginseng and persimmon vinegar have been proved in modern experiments as well as traditional textbooks.

Red ginseng is a red ginseng made by ginseng roots. Ginseng that can be used to make red ginseng is a perennial plant belonging to the genus Panax. ginseng), hwagisam (Panax quinquefolia), jeonchilsam (thirty-seven, Panax notoginseng), jukjeol three (Panax vietnamensis), Panax elegans tee climb (Panax elegatior), Panax Wan Jia Augustine (Panax wangianus) or Panax non pinra Tippi Douce (Panax bipinratifidus ) and Panax angustifolium . Red ginseng contains a lot of saponins. Most of the saponins in the plant family are oleanane and red ginseng saponins are the dammarane triterpenoid saponins that are rarely found in other plant systems. , Red ginseng saponin components include ginsenoside (ginsenoside) Rh ₂ , Rg ₂ , Rg ₃ , Rg ₁ , Rh ₁ . The general characteristics of saponins are found to be well soluble in water and alcohol, continuously foaming, physiologically detoxifying, and not toxic to overdose. In particular, ginsenosides are known to exert a variety of effects on the body regulatory function by affecting the central nervous system, endocrine system, immune system, metabolic system and the like. In addition, the vascular endothelium is known to promote the release of NO, which is known to relax blood vessels (Gen. Pharmac. 1995, 26 (3), 483-487). It is known that ginsenoside Rg ₃ exhibits the strongest activity. The ginsenoside Rg ₃ activates K ⁺ channels in vascular smooth muscle, thereby inhibiting the influx of intracellular Ca ²⁺ , thereby vascular relaxation. (Eur. J. Pharmacol. 1999, 367, 41-49). In addition, ginsenoside Rg ₃ is by increasing the generation of cGMP (cyclic guanosine monophosphate) in the body tissues in addition to facilitating the release of NO relaxation material of endothelial origin indicates a vasodilator acting vasoconstrictive factors on the other hand PGH ₂ ( Prostaglandin H ₂ ) by antagonistic action is understood to have a protective effect against circulatory disorders ('96 Korea-Japan Ginseng Symposium, 1990, 1 ~ 21).

Meanwhile, ginsenosides Ra ₁ , Ra ₂ , Ra ₃ , Rb ₁ , Rb ₂ , Rc, and Rd, which are protopanaxadiol (PPD) saponins, contain glucose, arabinose, and xyl on carbon 3 and 20. It is a glycoside combined with os, etc. It is a saponin group which accounts for about 70-80% of all saponins of red ginseng. Acid hydrolysis of these saponin groups with acetic acid produces a ginsenoside Rg ₃ as the sugar moiety bound to carbon 20 is released, and further, one molecule of glucose bound to carbon 3 of ginsenoside Rg ₃ is dropped to ginsenoside. It has been reported that Rh ₂ is generated (Korean Journal of Pharmacy, 1991, 35 (5), 432-437; Arch. Pharm. Res., 1995, 18 (3), 164-168; Korean Patent No. 78500)

In addition, in 1990, Okuda et al., Ehime University, Japan, reported that Arg-Fru-Glc (arginine-fructosyl-glucose) and Arg, which are ninhydrine-positive substances, are the unique bioactive substances of Korean red ginseng. Arginine derivatives belonging to the amino acid glycoside called -Fru (arginyl-fructose) have been isolated (Matsuura, Y., et al., Journal of traditional Medicine., 1994, 11 , 256-263). All of the arginine derivatives were generated by a maillard reaction during the heat treatment of red ginseng from fresh ginseng and reportedly produced from arginine and maltose contained in ginseng (Korean J. Ginseng Sci. 1996, 20, 389 ~ 415), Arg-Fru-Glc is contained about 5.4% in red ginseng (Hannam University Ph.D. dissertation, 2010), Arg-Fru digested by maltase in vivo, -Fru is known to mediate vasodilation by NO (藥用人蔘. 1995, 233 ~ 244).

Persimmon, on the other hand, is a ripe fruit of the persimmon tree ( Diospyros kaki ), which is used for food in autumn. On the one hand, the taste is characterized by sweet taste, cold nature, and no poison. Penis (歸經) acts on the heart, menopause, large intestine (大腸經). Lowers heat and moisturizes the lungs, quenching thirst, eliminating alcoholism and stopping diarrhea. Based on various raw material parts of persimmons, the Chinese herbal medicines are treated with medicinal herbs, roots, roots, leaves, leaves, seeds, and diseases. ), Thalamus, sipi, and sicily. Sicily, which is described in the herbaceous wood, is an edifice obtained by processing the immature fruit of persimmon. Take the blue, young and immature fruit, crush it, put it in the dock, pour a small amount of water, stir well, and stir several times. Leave it at room temperature for about 20 days, then squeeze the residue and leave the remaining colorless colloidal solution. This is sicily. Sicily has been reported to be effective in hypertension (Jung-Sup, et al., 2003, 915-918). In Japan, juice has traditionally been known to treat hypertension and suppress heart attacks (Chem. Pharm. Bull., 1990, 38, 1049-1052). Persimmon vinegar can be said to be manufactured similarly to the process of processing such as sicing and juice. Persimmon vinegar is rich in tannins and ascorbic acid (vitamin C), which lowers the acidity of foods, enhances its preservation capacity, stimulates the secretion of digestive fluids through sourness, and stimulates taste buds. It is known to restore fatigue quickly. To make persimmon vinegar simply, put persimmons in a colander or sieve, place a bowl under it, and cover it so that insects do not enter it. When this persimmon is put in a bottle or other container and sealed and aged for about a year, a vinegar with a rich taste is made. The longer the ripening period is, the blacker the taste becomes and the more the organic acid content is. Persimmon vinegar is effective when it is applied to athlete's foot or insect bites, and it has a refreshing taste after drinking greasy or instant foods.It also helps to recover from fatigue, prevent hangovers, relieve stress, sterilize, increase nutrients, and prevent adult diseases. It is known to be effective.

The inventors of the present invention while studying a method that can be used as a therapeutic agent for vascular diseases using the red ginseng extract and persimmon vinegar, when the mixture of red ginseng and persimmon vinegar and reacted according to the appropriate temperature and time, ginsenoside Rg ₃ And by increasing the content of Arg-Fru, and confirmed that the persimmon vinegar reaction mixture of the red ginseng extract can be used as a therapeutic agent for vascular diseases, the present invention could be completed.

Meanwhile, Korean Patent No. 759222 discloses a red ginseng vinegar beverage containing licorice, ogapi and rounde extract, red ginseng granules and persimmon vinegar and oligosaccharide, and Korean Patent No. 992800 to acid-treated fermented products of ginseng. Processed ginseng and extracts thereof having increased content of ginsenosides Rg ₃ and Rh ₂ are disclosed, and the Korean National Patent No. 635025 adds ginseng or ginseng extract to brewing vinegar and heat-extracts at 90-120 ° C. for 6-24 hours. Although ginseng extracts fortified with ginsenoside Rg ₃ have been disclosed, the preceding documents do not disclose that red ginseng or ginseng mixed with persimmon vinegar can be used as a therapeutic agent for vascular diseases. . In addition, although the antihypertensive effect of ginsenoside Rg ₃ is disclosed in Korean Patent No. 504966, a technique different from the composition of the present invention containing arginine derivative and other active ingredients of red ginseng extract in addition to ginsenoside Rg ₃ It can be said.

An object of the present invention is to provide a composition for the prevention or treatment of vascular diseases containing the persimmon vinegar reaction mixture of ginsenoside Rg ₃ and arginine derivatives enhanced red ginseng extract.

The present invention relates to a composition for preventing or treating vascular diseases containing red ginseng extract and persimmon vinegar. The vascular disease may be a disease selected from the group consisting of atherosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, heart failure, complications occurring after carotid angioplasty, cerebral infarction, cerebral hemorrhage, and stroke.

The red ginseng extract may be prepared using any extraction method commonly used, and for example, immersion (cold and warm), hot water extraction, ultrasonic extraction or reflux cooling extraction may be used, but is not limited thereto. In addition, the red ginseng extract may be prepared by extracting with an extraction solvent selected from conventional forms of water, aqueous alcohol solution, and alcohol, and these extracts may be prepared alone or by adding an acceptable herbal medicine. The alcohol may be a lower alcohol (C1 to C4 alcohol) having 1 to 4 carbon atoms, and may be preferably selected from ethanol, methanol, propanol, isopropanol and butanol.

Preferably, the red ginseng extract may be a red ginseng extract prepared by removing all the alcohol components from the 70-90% alcohol solution extract of red ginseng and concentrating the solid content to 60% or more.

In detail, the red ginseng extract is added to the red ginseng, 5-10 times the 70-90% alcohol solution of the weight of red ginseng, and then extracted by heating at 70-90 ℃ for 4-8 hours, the extract is filtered and filtered After obtaining, it may be a red ginseng extract prepared by removing all the alcohol components in the liquid and concentrated to 60% or more solid content.

In addition, the red ginseng extract was filtered and then added to the remaining red ginseng 5 to 10 times the 70 ~ 90% alcohol solution of the weight of the red ginseng and repeating the heat extraction process for 4 to 8 hours at 70 ~ 90 ℃ 1-5 times In addition, the filtrate of the extract extracted repeatedly can be used by concentrating with the first liquid obtained.

Persimmon vinegar reaction mixture of the red ginseng extract of the present invention, 5 to 20 times (preferably, 8 to 12 times) the persimmon vinegar of the weight of the red ginseng extract to the red ginseng extract and reacted for 1 to 24 hours at 70 ~ 90 ℃ It can manufacture. The reaction temperature is preferably 75 ~ 85 ℃ is the best, the reaction time is the best 12 ~ 18 hours. In addition, the reactant may be dried to prepare a powder, and for this purpose, a conventional drying method such as reduced pressure drying, spray drying, or freeze drying may be used.

The persimmon vinegar used in the present invention can be used commonly used persimmon vinegar, preferably a solid content of 2 to 4%, pH 3.0 to 3.5 can be used persimmon vinegar.

Therefore, the composition containing the red ginseng extract and persimmon vinegar of the present invention is preferably,

(Step 1) adding a red ginseng aqueous solution of 70-90% alcohol of 5-10 times the weight of red ginseng, heat-extracted at 70-90 ℃ for 4-8 hours to obtain a liquid phase by filtration;

(2 step) removing all the alcohol components of the filtration liquid and then concentrating so that the solid content is more than 60% to prepare a red ginseng extract;

(3 step) adding 5 ~ 20 times the persimmon vinegar of the red ginseng extract weight to the red ginseng extract and reacting at 70 ~ 90 ℃ for 1 to 24 hours; And,

(4 step) drying the reactant to prepare a powder;

It can be prepared to include.

In addition, the red ginseng extract is filtered in the first step, and the remaining red ginseng is added with an aqueous solution of 70 to 90% alcohol of 5 to 20 times the weight of red ginseng, and then heated and extracted at 70 to 90 ° C. for 4 to 8 hours. After repeated times, the filtered liquid of the extract extracted repeatedly can be used by concentrating with the liquid of the first step obtained first.

The concentration method of the two steps can be carried out using a conventional extract concentration process.

The pH of persimmon vinegar used in the above three steps is preferably in the range of 3.0 to 3.5, using a solid content of 2 to 4%.

In addition, if the heating time of the three process is out of 24 hours, ginsenoside Rg ₃ and Arg-Fru, which is an arginine derivative, may be decomposed and reduced.

Drying of the four steps may use a conventional drying method such as reduced pressure drying, spray drying, or freeze drying.

The composition containing the red ginseng extract and persimmon vinegar of the present invention prepared by the above method may bring an effect of enhancing ginsenoside Rg ₃ and Arg-Fru.

The extract content of the ginsenoside Rg ₃ and the arginine derivative-enhanced red ginseng extract and the active ingredient in the composition for the prevention or treatment of vascular diseases containing a composition containing persimmon vinegar is used in the form and purpose of use, the severity of the patient and the like. It may be appropriately adjusted according to the weight, based on the weight of the solid content may be 0.01 to 99.9% by weight, preferably 0.1 to 50% by weight, but is not limited thereto.

In addition, the composition may be administered in various oral and parenteral dosage forms during actual clinical administration, and when formulated, diluents or excipients such as fillers, weights, binders, wetting agents, disintegrating agents, and surfactants that are commonly used may be used. Can be used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which solid preparations contain at least one excipient such as starch, cellulose, carbonic acid in a composition containing the persimmon vinegar reaction mixture of red ginseng extract. Calcium, crospovidone, hard silicic anhydride, lactose, magnesium stearate, talc, enzyme treatment stevia, microcrystalline cellulose, magnesium stearate, gelatin and the like can be used in combination. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used.

The dosage of the composition containing the extract of the present invention may vary depending on the patient's worm, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of the disease, depending on the judgment of the doctor or pharmacist The dose may be divided once a day or several times at regular time intervals. For example, the daily dose may be 0.001 to 500 mg / kg, preferably 0.1 to 200 mg / kg, based on the active ingredient content.

In another aspect, the present invention provides a health functional food for the prevention or improvement of vascular diseases containing the persimmon vinegar reaction mixture of red ginseng extract. The health functional food may be various foods, beverages, food additives and the like.

The content of the extract as an active ingredient contained in the health functional food may be appropriately contained according to the form of the food and the intended use thereof. For example, the extract may be added in an amount of 0.01 to 50% by weight of the total food weight. It can be added at a ratio of 0.1 to 10% by weight. In addition, the health beverage composition may be added in a ratio of 0.02 to 10 g, preferably 0.1 to 5 g based on 100 ml.

In addition to containing the extract as an essential ingredient in the indicated ratio, the health beverage composition of the present invention is not particularly limited in the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose, for example polysaccharides such as maltose and sucrose, and conventional sugars such as dextrin and cyclodextrin. And sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. The proportion of natural carbohydrates is generally 1-20 g, preferably 5-12 g per 100 ml of the composition of the present invention.

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

In particular, red ginseng or persimmon, which is a raw material of the composition of the present invention, has been used in herbal medicine since ancient times. It was found to have no effect in vivo.

As described above, the composition of the present invention containing red ginseng extract and persimmon vinegar is excellent in increasing NO production and inhibiting angiotensin converting enzyme, such as atherosclerosis, hypertension, angina pectoris, myocardial infarction, ischemic heart disease, heart failure, and carotid angioplasty. It can be usefully used as a composition that can effectively prevent or inhibit vascular diseases, including postoperative complications, cerebral infarction, cerebral hemorrhage, and stroke.

1 is a flowchart illustrating a method of preparing a persimmon vinegar reaction mixture of the red ginseng extract disclosed in Example 2 of the present invention.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to ensure that the contents introduced herein are thorough and complete, and that the spirit of the present invention to those skilled in the art can fully convey.

Example 1 Preparation of Red Ginseng Extract

50 kg of 70% ethanol aqueous solution was added to 10 kg of red ginseng, and extracted by heating at 80 ° C. for 6 hours. The red ginseng extract was filtered and 50 kg of 70% alcohol solution was added to the remaining red ginseng and extracted by heating at 80 ° C. for 6 hours. The process was repeated once more, and the result was extracted by heating red ginseng at 80 ° C. for 6 hours. . In this way, the extracted extract was filtered through a 100μm filter to remove all the alcohol components, and concentrated to 60% solid content using a concentrator to prepare a 7.2kg red ginseng extract.

Example 2 Preparation of Persimmon Vinegar Reaction Mixture of Red Ginseng Extract

After mixing 100 g of red ginseng extract of Example 1 and 1200 g of persimmon vinegar, by reacting the red ginseng extract and persimmon vinegar for 1 to 24 hours at 70 ~ 90 ℃ temperature under the conditions of Table 1, to prepare a persimmon vinegar reaction mixture of red ginseng extract It was lyophilized to prepare a powder. Persimmon vinegar was used by purchasing a pH 3.0, 3% solids content.

Table 1

Condition	Temperature (℃)	Hour
Example 2-1	70	One
Example 2-2	70	3
Example 2-3	70	6
Example 2-4	70	12
Example 2-5	70	18
Example 2-6	70	24
Example 2-7	80	One
Example 2-8	80	3
Example 2-9	80	6
Example 2-10	80	12
Example 2-11	80	18
Example 2-12	80	24
Example 2-13	90	One
Example 2-14	90	3
Example 2-15	90	6
Example 2-16	90	12
Example 2-17	90	18
Example 2-18	90	24

Comparative Example 1. Preparation of Persimmon Vinegar Reaction Mixture of Comparative Red Ginseng Extract

Preparation of persimmon vinegar reaction mixture of the red ginseng extract in the same manner as in Example 2, but the temperature and time conditions were different under the conditions of Table 2.

TABLE 2

Condition	Temperature (℃)	Hour
Comparative Example 1-1	60	One
Comparative Example 1-2	60	3
Comparative Example 1-3	60	6
Comparative Example 1-4	60	12
Comparative Example 1-5	60	18
Comparative Example 1-6	60	24
Comparative Example 1-7	70	48
Comparative Example 1-8	80	48
Comparative Example 1-9	90	48
Comparative Example 1-10	100	One
Comparative Example 1-11	100	3
Comparative Example 1-12	100	6
Comparative Example 1-13	100	12
Comparative Example 1-14	100	18
Comparative Example 1-15	100	24

Comparative Example 2. Preparation of Persimmon Vinegar Reaction Mixture of Ginseng Extract

The ginseng extract was prepared in the same manner as in Example 1, and instead of the red ginseng extract, a persimmon vinegar reaction mixture was prepared under the conditions of Example 2-11.

Comparative Example 3 Mixture of Red Ginseng Extract and Persimmon Vinegar Not Warmed

100 g of red ginseng extract of Example 1 and 1200 g of persimmon vinegar were mixed and lyophilized without heating.

Experimental Example 1 Ginsenoside Content of Persimmon Vinegar Reaction Mixture of Red Ginseng ExtractQuantitative Analysis>

The reaction mixture of Example 2, Comparative Examples 1 and 2 and 5 g of the mixture of Comparative Example 3 were passed through an SEP-Pak column and eluted with methanol. The eluate was used as a YMC-Pack Pro C ₁₈ column (4.6 × 250 mm, 5 μm), and the mobile phase eluted acetonitrile and water in the ratio as shown in Table 3 (Waters 486 Tunable Absorbance Detector). The content of ginsenosides Rb ₁ and Rg ₃ was measured (standard of Rg ₃ : SML0184 of Sigma-Aldrich, standard of Rb ₁ : Sigma-Aldrich G0777), and analyzed three times for each sample, and the result of mean ± standard deviation. Was obtained and the results are shown in Table 4.

TABLE 3

Table 4

Condition	Rb ₁ (mg / g)	Rg ₃ (mg / g)
Example 2-1	11.234 ± 0.25	2.123 ± 0.11
Example 2-2	10.010 ± 0.45	3.730 ± 0.21
Example 2-3	9.560 ± 0.66	4.896 ± 0.12
Example 2-4	7.916 ± 0.55	6.573 ± 0.13
Example 2-5	6.276 ± 0.32	8.186 ± 0.04
Example 2-6	4.513 ± 0.27	8.943 ± 0.16
Example 2-7	9.345 ± 0.21	3.231 ± 0.24
Example 2-8	8.626 ± 0.79	5.876 ± 0.11
Example 2-9	6.273 ± 0.33	7.850 ± 0.11
Example 2-10	3.933 ± 0.23	9.590 ± 0.27
Example 2-11	2.020 ± 0.27	11.516 ± 0.60
Example 2-12	1.710 ± 0.22	10.793 ± 0.38
Example 2-13	6.342 ± 0.43	6.234 ± 0.21
Example 2-14	4.513 ± 0.50	8.866 ± 0.18
Example 2-15	2.333 ± 0.23	11.366 ± 0.71
Example 2-16	1.680 ± 0.69	9.976 ± 0.13
Example 2-17	0.636 ± 0.20	8.900 ± 0.04
Example 2-18	0.633 ± 0.92	8.160 ± 0.16
Comparative Example 1-1	13.345 ± 0.23	0.234 ± 0.01
Comparative Example 1-2	12.342 ± 0.12	0.332 ± 0.03
Comparative Example 1-3	12.634 ± 0.25	0.645 ± 0.02
Comparative Example 1-4	12.289 ± 0.33	0.912 ± 0.02
Comparative Example 1-5	11.296 ± 0.12	1.234 ± 0.04
Comparative Example 1-6	11.456 ± 0.24	1.543 ± 0.03
Comparative Example 1-7	2.223 ± 0.08	1.273 ± 0.30
Comparative Example 1-8	0.523 ± 0.20	2.960 ± 0.09
Comparative Example 1-9	0.166 ± 0.14	2.703 ± 0.16
Comparative Example 1-10	8.363 ± 0.04	3.533 ± 0.04
Comparative Example 1-11	2.254 ± 0.05	3.322 ± 0.05
Comparative Example 1-12	1.345 ± 0.03	2.241 ± 0.03
Comparative Example 1-13	1.433 ± 0.04	1.632 ± 0.04
Comparative Example 1-14	1.654 ± 0.02	1.233 ± 0.06
Comparative Example 1-15	1.745 ± 0.03	0.464 ± 0.07
Comparative Example 2	14.230 ± 0.52	0.221 ± 0.03
Comparative Example 3	12.130 ± 0.51	2.976 ± 0.08

As can be seen from Table 4, the ginsenoside Rg ₃ content was increased in the reaction mixture of Example 2 than the reaction mixture of Comparative Examples 1 and 2 or the mixture of Comparative Example 3, and the content of ginsenoside Rb ₁ was largely opposite. It was confirmed that the reduction.

Experimental Example 2. Confirmation of Arg-Fru-Glc and Arg-Fru Content of Persimmon Vinegar Reaction Mixture of Red Ginseng Extract

Arg-Fru-Glc and Arg-Fru content of the persimmon vinegar reaction mixture of the red ginseng extract of the present invention was confirmed (standards of Arg-Fru-Glc and Arg-Fru: Emboss Research Institute [Daejeon] products). To this end, 5 g of each of the reaction mixtures of Example 2, Comparative Examples 1 and 2 and the mixture of Comparative Example 3 were passed through an SEP-Pak column to remove the saponin component, and then the portion eluted in 15 ml of water was subjected to Bio-LC PAD assay. (The 10th International Symposium on Ginseng, 2010, using a CarboPac PA-1 column 4.0 × 250 mm, column temperature 30 ° C., injection volume 5 μl, flow rate 1.0 mL / min. The developing solvent was water and 250 mM sodium hydroxide (NaOH ) Was 50:50 (v / v) and measured using an Amperometric Detector (see Table 5 of analytical conditions), which is shown in Table 6.

Table 5

Detector (PAD)	Amperometric Detector
column	CarboPac PA-1 Column (4 × 250㎜)
Column temperature (℃)	30
Mobile phase	250 mM NaOH: DW = 50: 50
Flow rate	1.0ml / min
Injection volume	5 μl

Table 6

Condition	Arg-Fru-Glc (mg / g)	Arg-Fru (mg / g)
Example 2-1	8.945	9.384
Example 2-2	8.682	9.815
Example 2-3	7.865	8.701
Example 2-4	8.427	8.982
Example 2-5	8.137	7.751
Example 2-6	7.518	8.018
Example 2-7	7.652	8.540
Example 2-8	7.652	8.540
Example 2-9	7.963	8.324
Example 2-10	7.528	9.043
Example 2-11	4.350	10.384
Example 2-12	5.411	9.088
Example 2-13	9.234	8.993
Example 2-14	9.187	8.914
Example 2-15	6.353	8.756
Example 2-16	5.739	8.806
Example 2-17	4.012	8.405
Example 2-18	3.810	8.299
Comparative Example 1-1	13.344	1.446
Comparative Example 1-2	13.435	1.333
Comparative Example 1-3	13.554	2.424
Comparative Example 1-4	12.345	2.245
Comparative Example 1-5	12.434	2.434
Comparative Example 1-6	12.555	2.354
Comparative Example 1-7	11.234	2.063
Comparative Example 1-8	4.428	2.620
Comparative Example 1-9	2.901	1.720
Comparative Example 1-10	3.444	1.344
Comparative Example 1-11	3.433	1.435
Comparative Example 1-12	3.353	1.222
Comparative Example 1-13	3.544	1.343
Comparative Example 1-14	3.435	1.434
Comparative Example 1-15	3.345	1.522
Comparative Example 2	14.230	0.221
Comparative Example 3	12.130	2.976

Referring to Table 6, it can be seen that the content of Arg-Fru in the reaction mixture of Example 2 is higher than the reaction mixture of Comparative Examples 1 and 2 or the mixture of Comparative Example 3.

Experimental Example 3. Confirmation of Vascular NO Production of Persimmon Vinegar Reaction Mixture of Red Ginseng Extract

CPAE cells (calf pulmonary artery endothelial cells) were used to determine the effect of persimmon vinegar mixture of red ginseng extract on vascular NO production. CPAE cells were passaged in RPMI-1640 medium containing 10% FBS (fetal bovine serum) and penicillin-streptomycin (100 units / ml). To this end, CPAE cells were dispensed in a 12 well plate at a concentration of 5 × 10 ⁵ cells / well in 1 ml and incubated for 2 hours at 37 ° C. and 5% CO ₂ to attach cells, followed by removing the medium. 2, the reaction mixture of Comparative Examples 1 and 2 and 1 ml (1 mg / ml) of the medium containing the mixture of Comparative Example 3 were replaced, followed by incubation for 24 hours at 37 ° C. and 5% CO ₂ . Subsequently, 100 μl of the supernatant was added to a 96-well plate, and 100 μl of Griess reagent (1% sulfanilamide / 0.1% naphthylene diamine dihydrochloride / 2.5% H ₃ PO ₄ ) was added thereto, and left in a cow for 10 minutes. The absorbance at 540 nm was measured and is shown in Table 7. The concentration of NO ₂ ⁻ produced by the sample was determined by substituting the standard curve prepared using sodium nitrite. In addition, the amount of NO produced was expressed as a fold value when the non-warmed red ginseng extract of Comparative Example 3 and the persimmon vinegar mixture were set to 1.00.

TABLE 7

Condition	NO generation capacity (fold)
Example 2-1	2.10 ± 0.07
Example 2-2	2.21 ± 0.06
Example 2-3	2.21 ± 0.07
Example 2-4	2.41 ± 0.05
Example 2-5	2.51 ± 0.06
Example 2-6	2.38 ± 0.04
Example 2-7	2.28 ± 0.05
Example 2-8	2.16 ± 0.03
Example 2-9	2.21 ± 0.02
Example 2-10	2.25 ± 0.03
Example 2-11	2.46 ± 0.03
Example 2-12	2.33 ± 0.06
Example 2-13	2.22 ± 0.05
Example 2-14	2.33 ± 0.04
Example 2-15	2.64 ± 0.03
Example 2-16	2.35 ± 0.03
Example 2-17	2.46 ± 0.05
Example 2-18	2.33 ± 0.04
Comparative Example 1-1	1.32 ± 0.04
Comparative Example 1-2	1.43 ± 0.05
Comparative Example 1-3	1.32 ± 0.06
Comparative Example 1-4	1.44 ± 0.07
Comparative Example 1-5	1.35 ± 0.08
Comparative Example 1-6	1.33 ± 0.05
Comparative Example 1-7	1.42 ± 0.06
Comparative Example 1-8	1.36 ± 0.07
Comparative Example 1-9	1.27 ± 0.03
Comparative Example 1-10	1.35 ± 0.02
Comparative Example 1-11	1.46 ± 0.04
Comparative Example 1-12	1.35 ± 0.03
Comparative Example 1-13	1.24 ± 0.02
Comparative Example 1-14	1.35 ± 0.01
Comparative Example 1-15	1.23 ± 0.02
Comparative Example 2	0.33 ± 0.03
Comparative Example 3	1.00 ± 0.01

Referring to the results of Table 7, it can be seen that the reaction mixture of Example 2 was significantly increased NO production than the reaction mixture of Comparative Examples 1 and 2 or the mixture of Comparative Example 3.

In addition, the effects of NO production when the standard ginsenoside Rg ₃ and Arg-Fru are mixed and when the ginsenoside Rg ₃ and Arg-Fru are used alone (total concentration of the mixture and the composition alone) are the same. As a result, it was found that synergistic effect (more than 2.5 times) on NO-generating ability is observed when ginsenoside Rg ₃ and Arg-Fru are mixed than when ginsenoside Rg ₃ or Arg-Fru is alone. I could see that. Therefore, with reference to this, the persimmon vinegar reaction mixture of the red ginseng extract of Example 2 of the present invention was expected to increase the NO production ability due to the increased content of ginsenoside Rg ₃ and Arg-Fru and their synergistic effect.

Experimental Example 4. Confirmation of Inhibition of Angiotensin Converting Enzyme of Persimmon Vinegar Reaction Mixture of Red Ginseng Extract

Inhibitory activity of angiotensin converting enzyme was measured according to the method of Cushman and Cheung (1971) (Biochem Pharmacol., 1971, 20, 1637-1648). Thus, the reaction mixture of Example 2, Comparative Examples 1 and 2 and the mixture of Comparative Example 3 (sample solution, sample) 50 μl (1 mg / ml), HHL (25 mg hippuryl-L-histidyl-L-leucine / 2.5 150 ml of 0.1 ml sodium borate buffer (pH 8.3) solution and 100 µl of 0.1 M sodium borate buffer (pH 8.3) containing 0.4 M sodium chloride (NaCl) were mixed for 10 minutes at 37 ° C. incubation). To this was added 50 μl of coenzyme solution of angiotensin converting enzyme (supernatant recovered by centrifugation at 15,000 × g for 1 hour after stirring for 24 hours with 10-fold 0.1M borate buffer to ACE rabbit lung acetone powder). After the reaction for 30 minutes, 250 µl of 1N hydrochloric acid (HCl) was added to stop the reaction, and 1.5 ml of ethyl acetate was added thereto, followed by stirring for 15 seconds. Next, 1 ml of the supernatant obtained by centrifugation at 2,000 rpm for 20 minutes was completely dried at 100 ° C. for 40 minutes, and then 1 ml of distilled water was added and dissolved, and the absorbance of the sample was measured at 228 nm.

As a control, distilled water was used instead of the mixture solution. The blank sample (sample blank: HCl was used to stop the reaction first, and then the enzyme was measured for absorbance. The absorbance of the sample was subtracted from the absorbance of the sample. Cycle) 1N hydrochloric acid was added first, followed by addition of crude enzyme solution of angiotensin converting enzyme, 1.5 ml of ethyl acetate was added thereto, followed by stirring for 15 seconds, followed by centrifugation at 2,000 rpm for 20 minutes. Was taken, and completely dried at 100 ° C. for 40 minutes, and then dissolved in 1 ml of distilled water, and the absorbance was measured at 228 nm.

Inhibitory effect of the angiotensin converting enzyme according to the measured absorbance was calculated according to the following scheme and shown in Table 8. In addition, the reaction value was shown as the fold value when the mixture of the unheated red ginseng extract of Comparative Example 3 and persimmon vinegar was set to 1.00.

S: absorbance of sample solution

SB: Absorbance of blank sample (sample blank: enzyme added after stopping reaction by hydrochloric acid)

C: absorbance of the control group (control: buffer instead of sample)

B: Absorbance of the blank (Blank: HHL, sample, HCl, enzyme [coenzyme solution of angiotensin converting enzyme], ethyl acetate and the same process after)

Table 8

Condition	Inhibitory Effect of Angiotensin Converting Enzyme (fold)
Example 2-1	2.96 ± 0.03
Example 2-2	2.65 ± 0.04
Example 2-3	3.25 ± 0.05
Example 2-4	3.54 ± 0.05
Example 2-5	3.33 ± 0.08
Example 2-6	3.24 ± 0.07
Example 2-7	2.92 ± 0.06
Example 2-8	2.83 ± 0.06
Example 2-9	3.44 ± 0.04
Example 2-10	3.57 ± 0.03
Example 2-11	3.64 ± 0.02
Example 2-12	3.33 ± 0.05
Example 2-13	2.44 ± 0.04
Example 2-14	2.52 ± 0.03
Example 2-15	2.63 ± 0.02
Example 2-16	2.34 ± 0.05
Example 2-17	2.45 ± 0.04
Example 2-18	2.56 ± 0.03
Comparative Example 1-1	1.34 ± 0.05
Comparative Example 1-2	1.65 ± 0.06
Comparative Example 1-3	1.53 ± 0.05
Comparative Example 1-4	1.44 ± 0.04
Comparative Example 1-5	1.55 ± 0.06
Comparative Example 1-6	1.33 ± 0.05
Comparative Example 1-7	1.42 ± 0.08
Comparative Example 1-8	1.23 ± 0.07
Comparative Example 1-9	1.33 ± 0.02
Comparative Example 1-10	1.24 ± 0.03
Comparative Example 1-11	1.35 ± 0.02
Comparative Example 1-12	1.16 ± 0.04
Comparative Example 1-13	1.23 ± 0.03
Comparative Example 1-14	1.34 ± 0.02
Comparative Example 1-15	1.15 ± 0.03
Comparative Example 2	0.24 ± 0.03
Comparative Example 3	1.00 ± 0.06

Referring to the results of Table 8, it can be seen that the reaction mixture of Example 2 significantly increased the activity of angiotensin converting enzyme than the reaction mixtures of Comparative Examples 1 and 2 or the mixture of Comparative Example 3.

Experimental Example 5. Toxicity Test

5-1. Acute Toxicity

The present study was conducted to investigate the toxicity of the red ginseng extract mixture of the present invention to the animal body acutely (within 24 hours) when ingested in a short period of time and to determine the lethality. Twenty ICR mouse strains, which are common mice, were assigned to 10 control and experimental groups, respectively. In the control group, PEG-400 / tween-80 / ethanol (8/1/1, v / v / v) was administered only, and the experimental group prepared persimmon vinegar reaction mixture of red ginseng extract of Example 2-11 of the present invention. It was dissolved in / tween-80 / ethanol (8/1/1, v / v / v) and orally administered at a concentration of 2g / kg / day. As a result of investigating each mortality rate after 24 hours, it was confirmed that all mice survived in the control group and the experimental group administered the persimmon vinegar reaction mixture of 2g / kg / day red ginseng extract.

5-2. Long-term and tissue toxicity experiments in experimental and control groups

Long-term toxicity experiment was conducted by administering the persimmon vinegar reaction mixture of the red ginseng extract of the present invention to C57BL / 6J mice (10 per group) for 8 weeks at each concentration (final concentration 2g / kg / day). In order to investigate the effect on the organs (tissues) of the animal, the experimental group to which the persimmon vinegar reaction mixture of red ginseng extract of Example 2-11 of the present invention and PEG-400 / tween-80 / ethanol (8/1/1, Blood was collected after 8 weeks from animals in the control group that received only v / v / v), and blood concentrations of glutamate-pyruvate transferase (GPT) and blood urea nitrogen (BUN) were selected from the Select E (Vital Scientific NV, Netherland) instrument. Measured using. As a result, in the case of GPT, which is known to be related to hepatotoxicity, and BUN, which is known to be related to nephrotoxicity, there was no difference in the experimental group compared to the control group. In addition, liver and kidneys were excised from each animal, and histological observations were performed by optical microscopy through a conventional tissue fabrication process. No abnormalities were observed in all tissues.

Preparation Example 1. Pharmaceutical Formulations

1-1. Manufacture of tablets

200 g of the persimmon vinegar reaction mixture of the red ginseng extract of Example 2-11 of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silicic acid. 10% gelatin solution was added to the mixture, which was then ground and passed through a 14 mesh sieve. It was dried and the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into a tablet.

1-2. Preparation of Injection

1 g of the persimmon vinegar reaction mixture of the red ginseng extract of Example 2-11 of the present invention, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

Preparation Example 2 Food Preparation

2-1. Preparation of Cooking Seasonings

Persimmon vinegar reaction mixture of the red ginseng extract of Example 2-11 of the present invention was added to the cooking seasoning by 1% by weight to prepare a cooking seasoning for health promotion.

2-2. Manufacture of Flour Food

The persimmon vinegar reaction mixture of the red ginseng extract of Example 2-11 of the present invention was added to the flour at 0.1% by weight, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare health promoting foods.

2-3. Preparation of soups and gravy

The persimmon vinegar reaction mixture of the red ginseng extract of Example 2-11 of the present invention was added to the soup and the broth in 0.1 wt% to prepare a soup and broth for health promotion.

2-4. Manufacture of dairy products

Persimmon vinegar reaction mixture of red ginseng extract of Example 2-11 of the present invention was added to the milk at 0.1% by weight, and various dairy products such as butter and ice cream were prepared using the milk.

2-5. Vegetable Juice Manufacturing

0.5 g of persimmon vinegar reaction mixture of red ginseng extract of Example 2-11 of the present invention was added to 1,000 ml of tomato juice or carrot juice to prepare vegetable juice for health promotion.

2-6. Fruit juice manufacturing

0.1 g of the persimmon vinegar reaction mixture of the red ginseng extract of Example 2-11 of the present invention was added to 1,000 ml of apple juice or wine to prepare fruit juice for health promotion.

Claims

To the red ginseng extract prepared by concentrating the 1-4 carbon atoms of red ginseng or an aqueous solution of alcohol, red persimmon vinegar with pH 3.0-3.5 of 5-20 times the weight of red ginseng extract was added and reacted at 70-90 ° C. for 1-24 hours. A composition for preventing or treating hypertension, angina pectoris or myocardial infarction, characterized in that it contains a persimmon vinegar reaction mixture of the prepared red ginseng extract.
To the red ginseng extract prepared by concentrating the 1-4 carbon atoms of red ginseng or an aqueous solution of alcohol, red persimmon vinegar with pH 3.0-3.5 of 5-20 times the weight of red ginseng extract was added and reacted at 70-90 ° C. for 1-24 hours. Health functional food for the prevention or improvement of hypertension, angina pectoris or myocardial infarction, characterized in that it contains persimmon vinegar reaction mixture of the prepared red ginseng extract.
(Step 1) adding a red ginseng aqueous solution of 70-90% alcohol of 5-10 times the weight of red ginseng, heat-extracted at 70-90 ℃ for 4-8 hours to obtain a liquid phase by filtration;

(Step 2) removing all the alcohol or aqueous solution components of the filtration liquid and concentrated to a solid content of 60% or more to produce a red ginseng extract;

(Step 3) adding the persimmon vinegar of pH 3.0 ~ 3.5 of 5-20 times the weight of the red ginseng extract to the red ginseng extract and reacting for 1 to 24 hours at 70 ~ 90 ℃; And,

(4 step) drying the reactant to prepare a powder;

Method of producing a composition containing the persimmon vinegar reaction mixture of red ginseng extract, characterized in that it comprises a.