JP2008255075A - Improving agent for vessel endothelium function and health food - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a promoter for NO action, having no side effect from the view point of angiopathy, safe and continuously ingested, and to provide an improving agent and health foods for retaining and improving vascular endothelial cell functions and preventing arteriosclerosis, hypertension and diabetes. <P>SOLUTION: The vascular endothelium function improver contains a mixture of hyperoside and isoquercitrin as active ingredients. When both ingredients coexist, absorption into the body and metabolism are gently performed continuously so action and effects continue for a long time. As improvement of vascular endothelium functions, production of nitrogen monoxide and promotion of secretory function are listed and arteriosclerosis preventing function and hypertension preventing function are also improved. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a vascular endothelial function improving agent having a predetermined component as an active ingredient and a health food containing the same.

  Vascular endothelial cells are said to play an important role in maintaining homeostasis of the biological circulatory system by regulating vascular tonus and vascular permeability and functioning antithrombotic, that is, vascular endothelial cell damage is It has been revealed that it is deeply related to cardiovascular diseases such as hypertension and arteriosclerosis. Vascular endothelial cells are located in the innermost layer of the blood vessel and consist of a single cell layer.

  Recently, these vascular endothelial cells produce and secrete various physiologically active substances in addition to functioning as a barrier that separates the lumen of the blood vessel from the blood vessel wall. Particularly, nitric oxide (hereinafter abbreviated as NO) is very much. It is said that it is important.

  NO is considered as a factor that directly affects the above-described regulation of vascular tonus and vascular permeability, maintenance of blood pressure and insulin sensitivity, prevention of arteriosclerosis, and the like. It is also known that when NO production decreases due to hypertension, menopause, etc., vascular endothelial cell function is damaged, causing climacteric disorders such as arteriosclerosis (thickening of blood vessels), stiff shoulders, coldness, etc. .

  The mechanism by which vascular endothelial cells are damaged in vivo is not well understood, but the cause of these diseases or the treatment of arteriosclerosis, hypertension, diabetes, or Proposals have been made on drugs for preventing and treating vascular disorders caused by these. In particular, an agent for improving vascular endothelial cell function comprising fenofibrate as an active ingredient (Patent Document 1), an agent for preventing arteriosclerosis containing chlorogenic acid, caffeic acid or ferulic acid extracted from green coffee beans or southern sky leaves, Known are a NO action enhancer or a vascular endothelial function improving agent (Patent Document 2), and a vascular endothelial growth factor production promoter (Patent Document 3) composed of an extract of a plant of the genus Asagaoka. )

JP-A-5-194209 JP 2003-261444 A JP 2003-160503 A

  However, in the above-described prior art, the proposed vascular endothelial cell functional natural component has not been sufficiently enhanced because the material is still limited, and may be required more and more in the future. Drugs and health foods that can be safely and continuously ingested without any side effects in terms of vascular disorders with various ingredients and various ingredients for the prevention and treatment of highly arteriosclerosis (vascular thickening), hypertension, and diabetes Is required.

  Therefore, the object of the present invention is to solve the above-mentioned problems, and to find out that there is a new function by combining a new material and ingredients, and thereby there is no side effect from the aspect of vascular disorder, and it is safe and continuous ingestion. It is possible to provide a NO action enhancer, maintenance and improvement of vascular endothelial cell function, arteriosclerosis, hypertension or diabetes prevention agent useful for prevention of diabetes and health food.

  The inventors of the present application have conducted intensive research in view of the above-described present situation, and as a result, are highly safe so that they can be taken or ingested in the long term, have an effect of enhancing NO production, and have vascular disorders. The present invention has been completed by discovering a component that has preventive and ameliorating effects, and in particular, can achieve arteriosclerosis and prevention or treatment of hypertension or diabetes from the viewpoint of vascular disorders.

  That is, in order to solve the above-described problems, in the present invention, a vascular endothelial function improving agent containing a mixture of hyperoside and isoquercitrin as an active ingredient is provided. As used herein, improvement of vascular endothelial function includes promotion of NO production function and / or secretion function, and also includes improvement of arteriosclerosis prevention function or hypertension prevention function.

  The mixture of hyperoside and isoquercitrin may be a mixture of ingredients extracted from one or more plant bodies selected from Rafu, Hazama, Hawthorn, St. John's wort and Kaika. it can.

  In other words, hyperoside and isoquercitrin can be easily extracted and separated from plants such as leaves of Raffa, hawthorn, St. John's wort by ordinary chemical engineering extraction techniques using solvents such as water and alcohol, and can be purified. it can.

Raffma has long been drunk as tea in China, and its safety is well known. That is, it is preferable in terms of safety and availability of mixed components that the mixture of hyperoside and isoquercitrin is composed of Rahma's aqueous solvent extract.
Plants such as hawthorn and St. John's wort (also referred to as Hypericum perforatum) are also used as food and medicinal materials (herbs), respectively, and their safety is not a problem.

The action and effect of hyperoside and isoquercitrin were found to be particularly remarkable only when both coexist, and the present invention has been completed.
When hyperoside and isoquercitrin are each taken or ingested alone, the effects as seen in this invention are not exhibited. That is, the coexistence of hyperoside and isoquercitrin is a new finding and an essential condition of the present invention. Furthermore, when these two components coexist, they have also been found to have a feature that their effects are sustained for a long time because the absorption and metabolism in the body are carried out slowly and continuously.

  From these things, it can be set as the health food for vascular dysfunction prevention by setting it as the well-known food adjusted to contain the mixture of a hyperoside and isoquercitrin.

  As described above, the present inventors have found that the action on vascular endothelial cells has been discovered by the inventors of the present application, thereby preventing vascular disorders caused by hypertension, arteriosclerosis and diabetes, ie, vascular endothelial cell abnormalities. Drugs having safe and excellent preventive and therapeutic effects are presented, and are also effective in preventing and treating diseases such as hypertension, arteriosclerosis and diabetes.

  Since the present invention contains a mixture of hyperoside and isoquercitrin as an active ingredient, there is no side effect in terms of vascular disorders, it can be safely and continuously ingested, NO action is enhanced, and vascular endothelial cell function is improved. (Including functional retention), it has an effect of preventing arteriosclerosis and hypertension or diabetes, and therefore has an advantage that a health food can be presented by preparing a food containing the mixture.

The mixture of hyperoside and isoquercitrin used in the present invention contains approximately equal amounts of both components of a predetermined plant species, particularly in Rhuma, and therefore it is preferable to employ the plant body.
That is, the rough leaves contain 0.2% to 0.5% (dry basis) of both hyperoside and isoquercitrin. On the other hand, in hawthorn leaves, hyperoside is 1% and isoquercitrin is 0.1% (inventor's analysis example), and hyperoside is unevenly distributed.

  Kaika synthesizes isoquercitrin from rutin, an ingredient, and conversely, it becomes a raw material for isoquercitrin. Note that hyperoside and isoquercitrin can be easily analyzed using high-performance liquid chromatography (HPLC) of a general-purpose analytical instrument.

  The most suitable raw material is Raffaaceae, which has the scientific name Apocynum venetum L. and is a perennial herb that grows mostly in the northeast and northwestern parts of China. The active ingredient extracted from Raffma is not particularly limited in the part of the plant body, but it may be mainly collected from the leaves, and after the sun-dried materials are cut and dried, if necessary Is manufactured by hot water extraction and purification.

  The extract / purified product thus produced contains hyperoside and isoquercitrin in almost equal proportions as in Raffa leaves, and the product of the present invention is not required to adjust the proportion of these active ingredients again. It can be used as a raw material.

  In the case of using as a drug, the rough extract obtained by hot water extraction / purification is further highly purified by a generally known fractionation operation to obtain a drug containing a predetermined amount. On the other hand, in the case of health foods taken daily, it is not always necessary to purify the active ingredients to a high degree, tea-based beverages extracted with hot water, extract beverages produced by concentration, and concentrated and dried powder. It can be used as a body food.

  Raffma used in this invention can be used as a hot water extraction raw material only by drying, but especially when used as a health food material for tea / extract beverages and granular powders, the taste and taste are enhanced. It is preferable to roast.

  The active ingredients hyperoside and isoquercitrin can be extracted with hot water about 10 times as much as the raw material of Rumama, and extraction conditions of 90 ° C. to 100 ° C. and about 60 minutes are preferable. The extracted liquid is, for example, separated by a 150-mesh sieve and concentrated by a global concentrator under conditions of a temperature of 40 ° C. to 60 ° C. and a vacuum of 90 kpa to 99 kpa. The concentrate is subjected to a sterilization step after removing solids with a centrifuge, and then an extract is obtained.

  This extract is made into a tea-based beverage or an extract-based beverage by reduction with water, and is used as a raw material for producing high-purity hyperoside and isoquercitrin. Furthermore, this extract is dried in a spray dryer and granulated to become a powdery dried product. This dried product is used as a raw material for various products and drugs as in the case of the extract.

  The effects of hyperoside and isoquercitrin, which are the active ingredients of the drug and health food of the present invention, are the most effective and preferable efficacy when ingesting 40 mg (converted to 1 kg of rat body weight) per day under conditions where both components are present in substantially equal amounts. Is recognized. That is, the effective intake amount is 10 to 100 mg, preferably 30 to 50 mg.

  The effect on humans is that when a drug or food contains a functional ingredient and is ingested, it is generally more sensitive to animals than rats and other animals, so the required intake is generally small. . And it is well known that the degree is usually 1/50 to 1/100 of animal experimental examples, and it is converted to 1/50 for drugs and 1/100 for foods and drinks. Yes. Therefore, the amount of the above-mentioned active ingredients hyperoside and isoquercitrin is such that the daily intake for adults (60 kg in terms of body weight) is 12 mg to 120 mg, preferably 40 mg to 60 mg for drugs, and 6 mg to 60 mg for food and drink, preferably 20 mg to 30 mg.

  In the present invention, an agent for improving vascular endothelial function containing a mixture of hyperoside and isoquercitrin as an active ingredient is a tablet, granule, powder, capsule, infusion solution, hemodialysis solution, peritoneal dialysis by a known formulation technique. Arbitrary formulation forms such as liquids and solutions can be used, and an appropriate administration method can be selected and used depending on the purpose of use.

  Examples of components constituting the preparation of the present invention include excipients, binders, disintegrants, lubricants, coating agents, wetting agents, stabilizers, solubilizers, purified water, pH adjusters, preservatives. And can be prepared by a known and commonly used production method.

  Furthermore, the agent for improving vascular endothelial function of the present invention includes various kinds of noodles including cooked rice, miso, soy sauce, cup noodles, curry roux, stew, various soups, sausages, bamboo rings, rice cakes, hampened and other kneaded products, dressings, breads It can be added to beverages such as powdered milk, yogurt, milk beverages, lactic acid beverages, vegetable / fruit juice beverages, soft drinks, sports beverages, confectionery such as chocolate, cookies, chewing gum, and strawberries and used as health foods. Of course, it may be a health food in the above-described preparation form.

In the foods containing the vascular endothelial function improving agent of the present invention, dietary fiber, whey protein, whey protein hydrolyzate, saccharides, vitamin A, vitamin B1, ascorbic acid, vitamin D, vitamin E as necessary. Vitamins such as nicotinic acid amide, folic acid, calcium pantothenate, taurine, casein, lecithin, monoglyceride, polyglyceride, chlorophyll, etc. .
In addition, the vascular endothelial function improving agent of the present invention is added to animal feed such as pets as well as humans, and it is useful for improving various pathological conditions caused by vascular diseases such as hypertension as a health food for animals. It is.

  By the way, when the production amount of NO decreases due to lifestyle-related diseases, aging, etc., serious problems such as arteriosclerosis occur due to vascular endothelial cell dysfunction, arterial layer thickening / elasticity and lack of flexibility. It is well known to cause various diseases.

  The inventors of the present application conducted a study using this disease model system, a spontaneously hypertensive rat (SHR), and made the invention proposed here. The details will be described below.

  That is, a continuous oral administration test of SMC to each of hyperoside and isoquercitrin alone, a mixture of both components, and CMC (carboxymethylcellulose: food additive stabilizer) as a control substance was performed, and blood pressure was measured. As a result, no significant antihypertensive effect was observed with single oral administration of hyperoside and isoquercitrin.

  In contrast, a significant antihypertensive effect was observed when the hyperoside and isoquercitrin mixture was orally administered. Therefore, regarding SHR after continuous administration of a hyperoside and isoquercitrin mixture in which a significant antihypertensive action was observed, arterial vascular media thickness (morphometry), vascular endothelial NO synthase (eNOS) activity (immunohistological staining) The blood NO concentration (quantitative analysis) was evaluated and a tendency to suppress vascular thickening, an increase in eNOS activity, and a significant increase in blood NO concentration were observed.

  In addition, each of hyperoside and isoquercitrin alone and a mixture of both components was orally administered to SHR once, and blood metabolite concentrations were monitored for 24 hours immediately after administration. As a result, it was found that hyperoside was absorbed in the large intestine and isoquercitrin was absorbed in the small intestine. That is, it has been clarified that there is a time difference between the two components in absorption and metabolism in the body.

  When a mixture of hyperoside and isoquercitrin is administered, it is first observed that isoquercitrin is absorbed in the small intestine and later hypercoside is absorbed in the large intestine to maintain the quercetin metabolite concentration in the blood. It was. It is important that the vascular endothelium is exposed to these metabolites by the presence of a moderate concentration of quercetin metabolites in the blood, which circulate for a long time, as in the case of administration of a hyperoside and isoquercitrin mixture.

  As described above, the effect of the present invention occurs only when it is ingested and administered as a mixture of hyperoside and isoquercitrin, and the present invention has been made by elucidating a unique phenomenon through detailed examination. is there.

[Example 1]
SHR (♂, 5 weeks old) once a day, CMC (control), hyperoside [hereinafter abbreviated as HP. ] 40 mg / Kg (weight conversion), isoquercitrin [hereinafter abbreviated as IQ. ] 40 mg / Kg (weight conversion), hyperoside and isoquercitrin mixture [hereinafter abbreviated as HP + IQ. ] 40 mg / Kg (in terms of body weight) was continuously administered for 7 weeks (gastral forced oral administration), blood pressure was measured, and the results are shown in Table 1.
As is clear from the results in Table 1, no significant antihypertensive effect was observed in the [HP] and [IQ] single administration groups, but only in the [HP + IQ] administration group, compared with the control group. The effect was recognized.

[Example 2]
The total number of SHR (N = 7) in each of the control group shown in Example 1 and the 40 mg / Kg [HP + IQ] administration test group in which significant antihypertensive action was observed, Wistar Kyoto rats (WKY, N = For 3), the coronary artery of the heart was removed, a cross section of the blood vessel was prepared, and the thickness of the vascular media (smooth muscle) was measured by immunohistochemical staining.

FIG. 1 shows an example of a blood vessel cross-sectional photograph of a WKY normal control group, FIG. 2 an SHR control group, and FIG. 3 a 40 mg / Kg [HP + IQ] administration test group.
1 to 3, the portion dyed in a dark color in a ring (brown) indicates the vascular media. The thickness of the vascular media of WKY that does not cause an increase in blood pressure was thinner than SHR (control group) in which the blood pressure increased. That is, the thickness of the vascular media of SHR with increased blood pressure was significantly increased. And when SHR (control group) in which blood pressure increase was observed was compared with the vascular media thickness in which the increase in blood pressure was suppressed by [HP + IQ] administration, the latter tended to be thin.

Table 2 shows the results of statistical processing by measuring the thickness of 10 portions per leaf of one WKY / SHR blood vessel section.
A significant difference was observed between the WKY normal control group and the SHR control group, and in the SHR control group, after the blood pressure increased with time, thickening of the vascular media was observed. There was no significant difference between the SHR control group and the SHR [HP + IQ] administration test group due to the large variation, but the latter showed a tendency to become thin, that is, a tendency to suppress vascular media thickening was observed.

The total number of SHR (N = 7) in each of the control group shown in Example 1 and the 40 mg / Kg [HP + IQ] administration test group in which significant antihypertensive action was observed, WKY as a normal control group (N = 3) In the above, the heart coronary artery was removed, a blood vessel cross section was prepared, and the NO synthase (eNOS) activity of the vascular endothelium was observed by immunohistochemical staining.
Examples of the photographs are shown in FIGS. In these figures, the eNOS activity is indicated by the shade (dark brown) of the stained portion of the inner wall of the blood vessel. The higher the eNOS activity, the higher the degree of coloring and the dark brown color becomes darker.

  As is clear from FIGS. 4 to 6, the degree of coloring of the inner wall of WKY was the highest and the color was the darkest dark brown. In comparison, the coloring level was weak in the SHR control group. In the SHR [HP + IQ] administration test group, the degree of coloring was strong and brown. That is, it can be seen that the eNOS activity of SHR whose blood pressure increased over time showed a decreasing tendency, but the eNOS decreasing tendency was suppressed in the [HP + IQ] administration test group.

[Example 4]
For the total number of SHR (N = 7) in each of the control group shown in Example 1 and the 40 mg / Kg [HP + IQ] administration test group in which significant antihypertensive action was observed, blood was collected under anesthesia, The NO concentration ([NO ₂ + NO ₃ ] ion concentration) was measured. As is clear from the results shown in Table 3, there was a significant difference between the two, and it was found that the NO concentration in the [HP + IQ] administration test group was higher and the production amount of NO was higher than that in the control group with increased blood pressure. It was.

[Example 5]
CHR (control), 20 mg / Kg [HP], 20 mg / Kg [IQ], 40 mg / Kg [HP + IQ] were administered once to SHR (♂, 10 weeks old), followed by intragastric gavage. Blood was collected every 0.5, 1, 2, 6, 12, and 24 hours, and the concentrations of quercetin and isorhamnetin in each plasma were analyzed and quantified by LC-MS / MS method.

  Quercetin and isorhamnetin in plasma are metabolites when hyperoside and isoquercitrin are absorbed from the intestine and circulate in the blood, that is, substances corresponding to quercetin metabolites. The intermediate quercetin metabolite was enzymatically decomposed, and its aglycones quercetin and isorhamnetin were analyzed and quantified by LC-MS / MS method.

The results are shown in Table 4. What is extremely characteristic in the results of this test is the transition of blood concentrations of HP and IQ metabolites (quercetin and isorhamnetin). In HP, the metabolite maximum concentration (Cmax) is reached after 5 to 6 hours. Reached in 1-3 hours.
This means that HP is absorbed in the large intestine and IQ is absorbed in the small intestine, indicating that there is a time difference between the two components being absorbed and metabolized in the body. When [HP + IQ] was administered, Cmax was reached in 3 to 6 hours.

  In addition, the metabolite concentration in the blood at the time of [HP + IQ] mixture administration was significantly higher than that in the case of HP alone administration, and did not show a rapid increase in concentration as seen with IQ alone administration, and took 6 hours after administration. Showed a tendency to gradually increase and then gradually decrease. That is, when [HP + IQ] was administered, it became clear that a constant concentration of metabolites was maintained in the blood for a long time. Metabolite concentrations in blood when HP and IQ are administered alone are not continuous because they are divided into absorption in the large intestine and absorption in the small intestine. In intermittent administration such as once a day, blood vessels The action on the endothelium is not continuous but intermittent (in other words, pulsed).

Micrograph of blood vessel cross section of normal control group administered with WKY / CMC Micrograph of blood vessel cross section of SHR / CMC administration group Micrograph of blood vessel cross section of SHR • [HP + IQ] administration test group Micrograph of blood vessel cross section showing eNOS activity of vascular endothelium of WKY / CMC administration normal control group Micrograph of blood vessel cross section showing eNOS activity of vascular endothelium in SHR / CMC administration control group Micrograph of blood vessel cross section showing eNOS activity of vascular endothelium in SHR • [HP + IQ] administration test group

Claims (6)

  An agent for improving vascular endothelial function, comprising a mixture of hyperoside and isoquercitrin as an active ingredient.   The vascular endothelial function improving agent according to claim 1, wherein the improvement of the vascular endothelial function is promotion of nitric oxide production / secretion function.   The agent for improving vascular endothelial function according to claim 1, wherein the improvement of the vascular endothelial function is an improvement of an arteriosclerosis prevention function or a hypertension prevention function.   2. The mixture of hyperoside and isoquercitrin is a mixture of components extracted from one or more kinds of plants selected from Rafu, Rabama, Hawthorn, St. John's wort and Kaika. The agent for improving vascular endothelial function according to any one of -3.   The agent for improving vascular endothelial function according to any one of claims 1 to 3, wherein the mixture of hyperoside and isoquercitrin is an aqueous solvent extract of Rafuma.   A health food for preventing vascular dysfunction, comprising a mixture of hyperoside and isoquercitrin.