WO2006132223A1

WO2006132223A1 - Anti-hypertensive composition

Info

Publication number: WO2006132223A1
Application number: PCT/JP2006/311290
Authority: WO
Inventors: Masamitsu Moriwaki; Kazuhiro Emura
Original assignee: San-Ei Gen F.F.I., Inc.
Priority date: 2005-06-08
Filing date: 2006-06-06
Publication date: 2006-12-14

Abstract

Disclosed is a novel application of an enzymatically modified isoquercitrin as an active ingredient for an anti-hypertensive composition. The anti-hypertensive composition comprising an enzymatically modified isoquercitrin as an active ingredient can be used as a composition for oral administration, particularly a pharmaceutical composition or food composition which has an anti-hypertensive effect and is intended to be used for the prevention of the development of hypertension or the prevention of the progression of hypertension.

Description

Specification

Antihypertensive composition

Technical field

The present invention relates to an antihypertensive composition that is effectively used for preventing the development of hypertension or suppressing the development of hypertension. In particular, the present invention relates to a health food that is effectively used for preventing the development of hypertension and suppressing the development of hypertension.

Background art

[0002] Conventionally, rutin contained in buckwheat is said to have an action to improve hypertension, but it is known that it is based on the strengthening action of capillaries (for example, see Non-patent Document 1). . On the other hand, in recent years, it has been reported that taercetin and talysin, which are flavonoids like rutin, significantly reduce blood pressure in spontaneously hypertensive rats (Non-patent Documents 2 and 3). It has also been reported that flavonoids such as quercitrin and myricitrin have angiotensin converting enzyme inhibitory action and are useful as antihypertensive drugs (see Patent Document 1, etc.).

[0003] However, it has been reported that enzyme-treated isoquercitrin has a blood pressure increase-inhibiting action and an action to prevent the development of hypertension!

Non-Patent Literature 1: Vitamin Society of Japan, “Vitamin Handbook (2) Water-soluble vitamins”, 195-1 96 (1989)

Non-patent document 2: J. Duarte, et al, British Journal of Pharmacology 133, 117-124, 2001 Non-patent document 3: 1.C. Villar, et al., Planta Med. 68 (9), 847-850, 2002

Patent Document 1: JP-A-6-135830

Disclosure of the invention

Problems to be solved by the invention

[0004] An object of the present invention is to provide an antihypertensive composition capable of preventing the onset of hypertension by suppressing an increase in blood pressure for a person who tends to develop hypertension. The More specifically, the present invention provides an antihypertensive composition that has an excellent blood pressure elevation-inhibiting action and has high safety and can be continuously taken as, for example, a health food. For the purpose.

Means for solving the problem

[0005] The inventors of the present invention have conducted extensive research to solve the above-mentioned problems. As a result, an enzyme-treated isoquer citrin was applied to an animal (natural hypertensive rat) constructed so as to spontaneously develop hypertension. It was found that the occurrence of hypertension is significantly suppressed by oral administration, and that even if it is continuously administered, the safety is extremely high without adversely affecting the body. It was confirmed that isoquercitrin can be effectively used to prevent the development of hypertension and to suppress the development of hypertension. The present invention has been completed based on strong knowledge.

That is, the present invention has the following configuration:

Item 1. An antihypertensive composition comprising enzyme-treated isoquercitrin as an active ingredient.

[0007] Item 2. The antihypertensive composition according to Item 1, comprising an enzyme-treated isoquercitrin in an effective amount that exhibits an antihypertensive effect.

[0008] Item 3. The antihypertensive composition according to Item 1 or 2, which has an oral dosage form.

[0009] Item 4. The antihypertensive composition according to any one of Items 1 to 3, wherein the enzyme-treated isoquercitrin is contained at a rate of 1 mg to 10 g per daily dosage unit.

[0010] Item 5. The antihypertensive composition according to any one of Items 1 to 4, wherein the anti-hypertensive composition is formulated to contain 1 mg to 10 g of enzyme-treated isoquercitrin per daily dosage unit.

[0011] Item 6. The antihypertensive composition according to any one of Items 1 to 5, which is a pharmaceutical product.

[0012] Item 7. The antihypertensive composition according to any one of Items 1 to 5, which is a food.

[0013] Item 8. The antihypertensive composition according to item 7, wherein the antihypertensive composition is contained in a container, and the antihypertensive effect is described as an effect on the container or the package thereof.

Item 9. The antihypertensive composition according to Item 7 or 8, which is a food for specified health use for preventing the development of hypertension or suppressing the development of hypertension.

[0015] Item 10. A method for preventing the onset of hypertension or suppressing the development of hypertension, comprising administering an enzyme-treated isoquercitrin in an effective amount exhibiting an antihypertensive effect to a subject at risk of hypertension.

[0016] Item 11. Use of enzyme-treated isoquercitrin for the production of an antihypertensive composition. [0017] Here, "antihypertensive composition" is "blood pressure increase inhibitory composition", and "antihypertensive effect" and "antihypertensive effect" are "blood pressure increase inhibitory action" and "blood pressure increase inhibitory", respectively. It can also be referred to as “effect”.

The invention's effect

[0018] According to the composition comprising the enzyme-treated isoquercitrin of the present invention as an active ingredient, a person who tends to develop hypertension based on the antihypertensive action of the enzyme-treated isoquercitrin. Thus, it is possible to exert an effect of suppressing the rise of blood pressure and preventing the development of hypertension or the effect of suppressing the development of high blood pressure. Therefore, the composition of the present invention can be effectively used as a pharmaceutical composition (antihypertensive agent) intended or effective for preventing the development of hypertension or suppressing the development of hypertension. In addition, the composition of the present invention is effectively used as a health food for the purpose of preventing or preventing the development of hypertension or suppressing the development of hypertension (for example, including a specific health food that can clearly specify the use of such health). be able to.

[0019] Enzyme-treated isoquercitrin is a material conventionally used as a food additive. From this, it can be safely used as a component of the above-mentioned pharmaceutical or food

BEST MODE FOR CARRYING OUT THE INVENTION

[0020] The antihypertensive composition of the present invention comprises enzyme-treated isoquercitrin as an active ingredient.

[0021] Here, "enzyme-treated isoquercitrin" is obtained by acting a glycosyltransferase on isoquercitrin in the presence of a sugar donor, and is represented by the following formula: A mixture with a-glycosyl isoquercitrin to a degree dalcosylated.

[0022] [Chemical 1]

[0023] (wherein Glc represents a glucose residue, n represents 0 or an integer of 1 or more)

Specifically, in the above formula, the enzyme-treated isoquercitrin is composed of isoquercitrin in which the number of 4-course dalcose residues (η) is 0, and the number of glucose residues in the α-1,4 bond (η) force S It is a mixture with i or more, usually 1 to 15, preferably 1 to 10 a-glycosyl isoquercitrin.

[0024] The enzyme-treated isoquercitrin used in the present invention may be a mixture of various enzyme-treated isoquercitrins having different glucose group binding numbers (n). Even a kind of enzyme-treated isoquercitrin that is single! / ,.

[0025] Enzymatically treated isoquercitrin can be prepared by treating isoquercitrin with an enzyme having a glucose residue transfer action (hereinafter referred to as glucose residue transfer enzyme). Although not limited, in general, enzyme-treated isoquercitrin is glycosylated by transferring a glucose residue to isoquercitrin in an equimolar amount or more using a glucose residue transferase such as amylase, dalcosidase, or transdalcosidase. Can be manufactured.

[0026] The glucose source used in the case of glycosylation is not limited as long as it can be transferred to one molecule of S-isoquercitrin, which is one or more molecules of the glucose residue. For example, glucose, maltose, amylose Amylopectin, starch, starch liquefied product, starch syrup, cyclodextrin, and the like can be used. The amount of glucose source used is usually 0.1 to 20 parts by weight, preferably 0.5 to 15 parts by weight, more preferably 1 to: LO part by weight, based on 1 part by weight of isoquercitrin present in the reaction system. Can be mentioned.

[0027] Examples of glucose residue transferase include α-amylase (EC 3.2.1.1), α-darcosidase (EC 3.2.1.20), cyclodextrin glucanotransferase (EC 2.4.1.1 9) (hereinafter abbreviated as CGTase). Can be used.

[0028] CGTase is produced by bacteria such as Bacillus circulans, Bacillus macerans, Bacillus stearothermophilus, Bacillus megaterium, Bacillus polymixer and other genus Bacillus, and Klebsea genus such as Klebsiella pneumoniae. Any of these can be used freely in the present invention.

[0029] All of these glucose transferases are commercially available enzymes. A commercially available enzyme agent (for example, trade name: Contisym, manufactured by Amano Enzym Co., Ltd.) can also be used. The enzyme may be a crude product that is not necessarily purified. For example, the glucose residue transfer enzyme-producing bacterium is inoculated in a medium containing isoquercitrin, and the enzyme-treated isoquercitrin is produced by a reaction by fermentation. A transferase or glucose residue transferase-producing bacterium may be immobilized and reacted with isoquercitrin in a batch or continuous manner to produce enzyme-treated isoquercitrin. Glucose residue transferase can be performed using each of amylase, darcosidase, or transdalcosidase alone, or can be used in combination (simultaneously or successively).

[0030] The reaction conditions for the glucose residue transferase may be any conditions under which the glucose residue transferase acts in a mixed water system of isoquercitrin, glucose residue transferase and the above glucose source. The amount of glucose residue transferase used is 1 part by weight of isoquercitrin, and when the glucose residue transferase is CGTase (enzyme specific activity is about 100 units (generates lmg of j8-cyclodextrin per minute from soluble starch) Enzyme amount is 1 unit)] It can be appropriately selected from the range of 0.001 to 20 parts by weight. Preferably, it is about 0.05 to about LO parts by weight, more preferably about 0.01 to 5 parts by weight.

[0031] The amount of isoquercitrin in the reaction system is not particularly limited, but for the purpose of efficiently carrying out glycosylation, usually 0.1 to 30% by weight, preferably 100% by weight in the reaction system. Is preferably contained in a proportion of 0.5 to 20% by weight, more preferably 1 to 10% by weight.

[0032] The temperature of the reaction system can be selected by appropriately selecting a range of about 80 ° C or less, which varies depending on the type of enzyme used. Within this range, industrially advantageous is about 20-80 ° C, preferably about 40-75 ° C. The pH condition is usually pH 3 to: about L 1 or less, preferably pH 4 to 8.

[0033] The reaction can be carried out while standing or stirring or shaking. In order to prevent oxidation during the reaction, it is also possible to add an anti-oxidation agent such as ascorbic acid that replaces the head space of the reaction system with an inert gas such as nitrogen. is there.

[0034] Thus, the glucose group binds to the glucose residue of isoquercitrin to produce the target enzyme-treated isoquercitrin. [0035] The number of glucose groups bound to the glucose residue of isoquercitrin (the number of n in the above formula (1)) is not particularly limited, but is usually 1 to 15, preferably 1 as described above. ~: Can be arbitrarily adjusted to be in the LO range. As a powerful adjustment method, for example, after the production of enzyme-treated isoquercitrin, various amylases (a-amylase, / 3-amylase, darcoamylase, di-darcosidase, maltase, etc.) are treated alone or in combination. Can be mentioned. By carrying out like this, the number of glucose sugar chains in the enzyme-treated isoquercitrin molecule obtained by the above-mentioned method can be decreased to obtain an enzyme-treated isoquercitrin having an arbitrary glucose sugar chain length.

[0036] The method for isolating and purifying the enzyme-treated isoquercitrin is not particularly limited. For example, as an isolation method, there can be mentioned a method of isolation using gel filtration resin in a conventional manner. The purification of the enzyme-treated isoquercitrin is not particularly limited, and can be carried out by arbitrarily combining conventional methods. Specifically, various resin treatment methods (adsorption method, ion exchange method, gel filtration method, etc.), membrane treatment method (ultrafiltration membrane treatment method, reverse osmosis membrane treatment method, ion exchange membrane treatment method, zeta potential membrane) Treatment method, etc.), electrodialysis method, salting out, acid precipitation, recrystallization, solvent fractionation method, activated carbon treatment method and the like.

[0037] The enzyme-treated isoquercitrin thus obtained is isoquercitrin (quercetin 3-0).

A further equimolar amount of glucose was bound to the glucose residue of monodalcoside)

-It is based on glycosyl isoquercitrin and is readily soluble in water.

[0038] As shown in the following test examples, enzyme-treated isoquercitrin was orally administered to rats (natural hypertensive rats) constructed to spontaneously develop hypertension with age. It was confirmed that the increase in blood pressure accompanying aging was suppressed for both systolic blood pressure, mean blood pressure and diastolic blood pressure. Furthermore, it was confirmed that the enzyme-treated isoquercitrin is extremely safe without adversely affecting the health of the rats even when continuously administered orally.

[0039] From these results, enzyme-treated isoquercitrin is effective in preventing the onset of hypertension in warm-blooded animals containing humans with a tendency to develop hypertension by suppressing an increase in blood pressure. Since it is possible to suppress the development of blood pressure, it is effective as an active ingredient in antihypertensive agents, antihypertensive agents, or antihypertensive agents, and for the effective production of health foods with powerful effects As a minute, it can be used effectively.

[0040] It should be noted that hypertension or hypertension means a state in which systolic blood pressure is 140 mmHg or higher or diastolic blood pressure is 90 mmHg or higher according to WHO criteria. Hypertension or hypertension targeted by the present invention is not limited, but preferably high blood pressure itself is the true form of this disease, and essential hypertension in which blood pressure increases with aging (essential hypertension) Symptom).

[0041] In addition, enzyme-treated isoquercitrin has the action of preventing the development of hypertension by suppressing an increase in blood pressure, or the progression of hypertension, so that it develops a disease caused by hypertension (hypertensive It can be used effectively for prevention of disease). Possible hypertensive diseases include, for example, hypertensive heart hypertrophy, hypertensive heart disease, hypertensive erythrocytosis (Gais-Beck syndrome), hypertensive retinopathy, hypertensive nephrosclerosis, hypertensive arteriosclerosis, hypertensive Examples include cerebral hemorrhage and hypertensive diencephalon syndrome (Page syndrome).

[0042] Therefore, according to the present invention, an antihypertensive composition comprising enzyme-treated isoquercitrin as an active ingredient can be provided.

[0043] In the present invention, the "antihypertensive composition" has an action of suppressing an increase in blood pressure, and is effective for preventing the development of hypertension, suppressing the development of hypertension, or preventing the development of a hypertensive disease caused by hypertension. It means a composition that can be used for More preferably, it is a composition that has anti-hypertension, in particular, prevention of the development of hypertension, or suppression of the development of hypertension, and is used for the specific health use. Examples of powerful compositions include pharmaceutical compositions, special-purpose foods such as foods for the elderly and foods for the elderly, functional foods such as nutritional functional foods and foods for specified health use, and similar health foods.

[0044] Specified health foods (including conditional specified health foods, standard-standard specified health foods and disease risk reduction labeled specified health foods) are approved by the Ministry of Health, Labor and Welfare in their packaging containers, etc. It is a food that can be labeled for approved functions or health use. However, the present invention is not limited to this, and any person may be provided with an indication that a person who takes the food for a specific health purpose can expect the purpose of the health through the intake. In the present invention, a specific health purpose is anti-hypertension, specifically, prevention of the development of hypertension (preferably essential hypertension) or suppression of the development of hypertension. To "suppress blood pressure rise", "prevent hypertension", "maintain blood pressure normally", "if you are concerned about blood pressure", "if you want to increase blood pressure", etc. Can do. Because foods for specified health use can display such functions, they can be differentiated from general foods that cannot display functions. / This is a preferred embodiment of food.

[0045] In the preparation of a pharmaceutical composition, enzyme-treated isoquercitrin is dissolved in water, alcohol (eg, ethanol), or other solvent in a solution state (emulsion, liquid, syrup, etc.) ) Or a solid state (powder, powder, granule, tablet, pill, capsule, chewable, etc.) prepared or molded by a known method and having various oral dosage forms can do.

[0046] In addition to the enzyme-treated isoquercitrin, these preparations may contain a pharmaceutically acceptable carrier or additive depending on the product form. For example, excipients for preparing solid-state formulations include lactose, sucrose, glucose, corn starch, gelatin, starch, dextrin, calcium phosphate, calcium carbonate, synthetic and natural aluminum silicates, magnesium oxide, dry water Examples include acid aluminum, magnesium stearate, sodium bicarbonate, and dry yeast. In addition, examples of excipients for preparing a preparation in a solution state include water, glycerin, propylene glycol, simple syrup, ethanol, ethylene glycol, polyethylene glycol, sorbitol and the like. In addition, these preparations are optionally formulated with a stabilizing agent such as citrate, phosphoric acid, malic acid or salts thereof; a sweetener such as sucralose or acesulfame potassium; a sweetener such as sucrose or fructose; alcohol , Preservatives such as glycerin; conventional or other suitable methods that may be mixed with conventional additives such as demulcents, diluents, buffers, flavors and colorants, powders, It can be produced into a preparation for oral administration such as tablet, emulsion, capsule, granule, chewable, liquid, or syrup.

[0047] The anti-hypertensive composition in a potent pharmaceutical form is preferably formulated to contain enzyme-treated isoquercitrin in the range of lmg to: LOg per daily dosage unit. Such a dose can be appropriately adjusted according to various factors such as the health condition of the person taking the enzyme-treated isoquercitrin, the administration method, and combinations with other agents (or food ingredients). It is preferably 1 mg to 5 g, more preferably 1 mg to 2 g. [0048] In addition, the enzyme-treated isoquercitrin described above can prevent the onset of hypertension by suppressing an increase in blood pressure in warm-blooded animals including humans who are prone to develop hypertension. Is also useful as an active ingredient in health foods used to suppress the development of hypertension.

[0049] Here, health food refers to food with the purpose of health, health maintenance and promotion in a more positive sense than ordinary food. The enzyme-treated isoquercitrin of the present invention is prepared in the form of a preparation as described above using a food-acceptable carrier or additive (powder, tablet, emulsion, capsule, granule, chewable, liquid, syrup, etc. ) And can be provided as a supplement for the prevention, treatment or amelioration of hypertension, or added to general food (in other words, used as one of the ingredients of food) Therefore, it can be used to prepare health foods (for example, functional foods, foods for specified health use) with the objective and efficacy of preventing the development of hypertension or suppressing the development of hypertension.

[0050] The type of food is not particularly limited. (1) Milk drink, lactic acid bacteria drink, soft drink with fruit juice, soft drink, carbonated drink, fruit juice drink, vegetable drink, vegetable, fruit drink, powdered drink, coffee Beverages, tea drinks, tea drinks, milk drinks, soy milk drinks, cocoa, sports drinks, supplement drinks, green juice drinks, etc .; (2) Puddings such as custard pudding, milk pudding, souffle pudding, fruit juice pudding, etc. Desserts such as jelly and bavaroa; (3) Ice cream, ice milk, lact ice, milk ice cream, ice cream and soft ice cream with fruit juice, ice candy, sorbet, ice confectionery, etc .; (4) chewing gum and balloons Gum and other gums (plate gum, sugar-coated grain gum); (5) Coated chocolate such as plate chocolate and marble chocolate, strawberry Chocolate such as chocolate, blueberry chocolate, melon chocolate and other flavored chocolates; (6) Hard candy (including bonbon, butterball, marble, etc.), soft candy (caramel, nougat, gummy candies, marshmallows, etc.) (7) Hard biscuits, cookie, baked confectionery such as rice crackers, rice crackers, etc. (The above (2) to) are collectively called confectionery); (8) Ketchup Seasonings such as sauce, soy sauce, dressing, miso, sugar, salt, mayonnaise, vinegar, black vinegar; (9) Livestock products such as ham, sausage, bacon, frozen hamburger; (10) Mouth spray Medicine, troches, swallowing supplements, drinks, granules, powders, tablets, etc. Quasi-drugs, health supplements; (11) pastes such as marmalade, jam, margarine, butter, hula paste, peanut paste; (12) fish ham, fish sausage, salmon salmon, chikuwa, hampen, tempura, etc. (13) Retort curry, retort soup, retort stew and other retort products; (14) Udon, buckwheat, Chinese soba, spaghetti, macaroni, dried salmon, somen, instant ramen ; (15) Alcoholic beverages such as fruit wine, liqueur, chuhai, carbonated alcoholic beverages such as red wine; (16) Soybean processed foods such as tofu and fried chicken; (17) Various fermentations such as pickles, cheese, yogurt, tempeh, and natto Foods: (18) Examples include various pet foods such as dog food and cat food.

[0051] Above all, from the viewpoint of continuous taking, confectionery such as bread, candy and tablet confectionery, other solid products, liquid products such as soft drinks and nutritional beverages, and semi-solid products such as jelly. It can illustrate suitably. More preferably, it is a beverage.

[0052] It is preferred that the anti-hypertensive composition in the form of a strong food is prepared to contain enzyme-treated isoquercitrin in the range of lmg to 10g per daily dose U. Such a dose can be appropriately adjusted according to various factors such as the health condition of the person taking the enzyme-treated isoquercitrin, the administration method, and the combination with other agents (or food ingredients). It is preferably 1 mg to 5 g, more preferably 1 mg to 2 g.

[0053] Especially when it is prepared in the form of a beverage, there is no restriction! However, the amount of enzyme-treated isoquercitrin is lmg to 10g, preferably lmg to 5g, more preferably lmg to 2g per 500mL volume of beverage. It is preferable to add enzyme-treated isoquercitrin so that the ratio of Example

[0054] Preparation examples, test examples, and examples are described below in order to clarify the constitution and effects of the present invention. However, the present invention is not affected by these examples.

[0055] mmm

(1) Preparation of isoquercitrin

After immersing 250 g of bud of Enju, a leguminous plant, in 2500 mL of hot water (above 95 ° C) for 2 hours, the filtrate obtained by filtration was obtained as the “first extract”. Meanwhile, the filtered residue is further heated. Extraction was performed by immersion in water to obtain a “second extract”. These first and second extracts are combined, cooled to 30 ° C or lower, the precipitated components are filtered off, and the precipitate is washed with water, recrystallized and dried to obtain 22.8 g of rutin with a purity of 95% or more. Got.

[0056] 20 g of this rutin was dispersed in 400 ml of water and adjusted to pH 4.9 using a pH adjuster. To this was added 0.12 g of Naringinase (Amano Enzym Co., Ltd., trade name Naringinase “Amano”, 3,000 U / g) to start the reaction, which was maintained at 72 ° C. for 24 hours. Thereafter, the reaction solution was cooled to 20 ° C., and the precipitate generated by cooling was separated by filtration. The obtained precipitate (solid content) was washed with water and dried to collect 13.4 g of isoquercitrin (hereinafter referred to as “IQC”).

[0057] (2) Preparation of enzyme-treated isoquercitrin

The IQClOg obtained above was dispersed with 500 mL of water and 40 g of corn starch. The reaction was started by adding 15 g of cyclodextrin glucanotransferase (CGTase: Amano Enzyme Co., Ltd., trade name Contizim, 600 UZml), and this was maintained for 24 hours under the conditions of pH 7.25 and 60 ° C. . After cooling the obtained reaction liquid, it was added to a column (Φ 3.0 X 40 cm) of Diaion HP-20 (manufactured by Mitsubishi Kasei Kogyo Co., Ltd.) and washed with lOOOmL of water. Next, 600 mL of 50% ethanol aqueous solution was applied to the column, and the resulting eluate was concentrated under reduced pressure and then lyophilized to obtain 12.8 g of enzyme-treated isoquercitrin (hereinafter referred to as “EMIQ”). .

[0058] The resulting EMIQ, and subjected to HPLC analysis under the following conditions, various α contained in EMIQ - calculated glycosylation molar ratio Louis Suk El citrine and _(0/0).

[0059] <HPLC conditions>

Column: Inertsil ODS— 2 Φ 4.6 X 250mm (GL Science)

Eluent: Water Z-acetonitrile ZTFA = 850Z150Z2

Detection: Absorbance measurement at a wavelength of 35 lnm

Flow velocity: 0.8m min.

[0060] The molar composition ratio (%) of EMIQ is shown below. The molar composition ratio is a composition ratio converted from IQC to IQC, where the total of eight components from IQC + Glc7 in which seven glucoses are bonded by α-1,4 bonds is defined as 100%. In addition to these components, EMIQ contains a trace amount of 8 or more glucoses bound to IQC (IQC + Glc8 or more). [0061] [Table 1]

IQC: Isoquercitrin Glcl Gl c7: Number of glucose additions to IQC

[0062] (3) Preparation of Taercetin (QC)

The IQClOg obtained in Preparation Example 1 was charged with 500 mL of 2N aqueous sulfuric acid and hydrolyzed in a boiling water bath for 2 hours. The obtained reaction solution was cooled to 30 ° C. or lower and then filtered to obtain crude quercetin. The obtained crude quercetin was washed with water, recrystallized and dried to recover 5 g of taercetin (hereinafter “QC” t) having a purity of 95% or more.

[0063] Test Example 1 Effect of EMIQ on blood pressure

Preparation Example 1 EMIQ prepared in (2) was orally administered to 5-week-old spontaneously hypertensive rats twice a day for 5 weeks to examine the inhibitory effect on blood pressure rise, mainly using changes in systolic blood pressure as an index. did. In parallel with this, as a comparative test (positive control), blood pressure increased by administration of diltiazem (diltiazem hydrochloride, manufactured by Wako Pure Chemical Industries, Ltd.), a calcium antagonist with antihypertensive action. Observed inhibitory effect (Hiroshi Narita, Taku Nagao, Masanori Inamasu, Hitoshi Iwasaki, Takashi Morita: Effects of Diltiazem on SHR blood pressure increase and associated cardiac hypertrophy and vascular thickening, Journal of Japanese Pharmacology 86 (3) 165-174 1985).

[0064] 1. Test animals and rearing conditions

In the test, male SHR / Izm rats (SPF, Japan SLC Co., Ltd.), which are commonly used in pharmacological studies as spontaneously hypertensive rats, were used. Animals were obtained at 4 weeks of age (body weight 45 70 g), with a quarantine period of 5 days followed by a acclimation period of 34 days, during which animals that were not abnormal were used for the study. The body weight range at the start of administration was 85 116 g.

[0065] Animal temperature 20 26 ° C (actual value: temperature 20 25 ° C), humidity 40 70% (actual value: humidity 40 61%), light and dark 12 hours each (lighting: 6am to 6pm) The animals were kept in an animal breeding room maintained at a ventilation rate of 12 times Z (fresh air sterilized by a filter). Blood pressure was measured in a soundproof room.

[0066] Animals are individually raised using stainless steel five-cage cages (W: 755 XD: 210 XH: 170 mm). did. The feed was a solid feed (CRF_1, Oriental Yeast Co., Ltd.) within 5 months after manufacture, and tap water was freely consumed.

[0067] During the test, the test animals were divided into 3 groups (each group, 10 animals): a control group (administration with water for injection), an EMIQ administration group, and a diltiazem administration group (positive control group). In addition, after dividing the weight and systolic blood pressure into layers using a computer, the average weight and average systolic blood pressure of each group and their variances were approximately equal by random sampling. On the day of administration start.

[0068] 2. Test method

(1) Administered sample and its administration method

The EMIQ prepared by the method described in the above preparation example was used. In addition, diltiazem hydrochloride (manufactured by Wako Pure Chemical Industries, Ltd.) was used as the positive reference substance diltiazem. These samples were used after being dissolved in water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.) at a predetermined concentration after weighing the required amount. In the control group, this water for injection (manufactured by Otsuka Pharmaceutical Factory) was used as the administration solution.

[0069] The number of doses was twice daily, in the morning and afternoon, and each dose was closest to the date of administration! Based on body weight, the EMIQ was 13 mg / kg / dose per day (daily dose) , 26 mg / kg / day), diltiazem is Yoshida et al. (Masumi Yoshida et al .: Medicinal pharmacological study of medicinal herb preparations. And Narita et al. (Hiroshi Narita et al .: Effects of Diltiazem on SHR blood pressure elevation and associated cardiac hypertrophy and vascular thickening, and reports on the effect of blood pressure elevation based on the report of Japanese Pharmacological Journal 86 (3): 165-174, 1985) 6 Omg / kg / dose (daily dose, 120 mg / kg / day). Also. Administration was oral, and oral gavage was performed using a polypropylene disposable syringe fitted with an oral sonde for rats.

[0070] (2) General condition observation and body weight measurement

For each test animal (10 animals per group) in each group (control group, EMIQ administration group, diltiazem administration group), administration day 1 (grouping day), 8, 15, 22, 29 and 36 days Prior to morning administration, general conditions were observed and body weights were measured.

[0071] (3) Measurement of blood pressure and heart rate For each test animal (10 animals in each group) in each group (control group, EMIQ administration group, diltiazem administration group), measure blood pressure before administration and on days 8, 15, 22, 29 and 36 Went. The blood pressure during the administration period was measured from about 1 hour after the morning administration to before the afternoon administration. For blood pressure, tail arteriovenous pressure was measured using a non-invasive automatic blood pressure measuring device (BP-98A, Softlon Co., Ltd.) and under anesthesia, tail caff method, systolic blood pressure, average blood pressure, diastolic blood pressure And heart rate was measured. The measurement was performed three times, and the average value was taken as the measurement value for each individual. Prior to blood pressure measurement, the animals were placed in a heat insulation box (set temperature: 38 ° C) for about 5 minutes in order to raise the body temperature of the animals.

[0072] For the numerical values obtained in the above experiment, the average value and standard deviation were calculated in each test group (control group, EMIQ administration group). Significance test was performed by F-test for equal variance. For equal variance, Student's t-test, and for unequal variance, Aspin-Welch's t-test. The significance level was 20% for the F test and less than 5% for other cases, and was divided into less than 5% (p <0.05) and less than 1% (p <0.01).

[0073] 3. Test results

(1) General condition and weight

Figure 1 shows the changes in body weight of each group (control group, EMIQ administration group, diltiazem administration group). As can be seen from the figure, during the administration period, there was no change in general condition or change in body weight that should be noted in any administration group, and there was a smooth change in body weight.

[0074] (2) Blood pressure and heart rate

2.1. Systolic blood pressure

Figure 2 shows the measurement results of systolic blood pressure. As can be seen from the figure, the mean value of systolic blood pressure in the control group (101) showed 136 mmHg before the start of administration, but subsequently increased with aging and showed 208 mmHg on the 36th day of administration. It was. In the EMIQ administration group (dose-), a significantly lower value (p <0.01, Aspin-Whlch's t-test) was observed compared to the control group on the 22nd and 36th days of administration, and the increase was significantly suppressed. . In addition, the positive control substance diltiazem administration group (1 △ 1) also had a significantly lower value compared to the control group on the 22nd, 29th, and 36th day (p 0.01, Aspin-Whlch's t-test). Was observed and the increase was suppressed. The inhibitory effect of the diltiazem-administered group was stronger than that of the EM IQ-administered group and after 22 days, a significantly lower value than the EMIQ-administered group (p <0. 05, Student's t-test) was accepted.

[0075] 2.2. Mean blood pressure and diastolic blood pressure

The measurement results of mean blood pressure are shown in Fig. 3, and the measurement results of diastolic blood pressure are shown in Fig. 4. As can be seen from Fig. 3, the mean blood pressure of the control group (-〇-) showed 108 mmHg before the start of administration, but then increased with aging, and showed 178 mmHg on the 36th day of administration. . The EMIQ administration group (sip-) showed a low value compared to the control group, and suppressed the increase. In addition, the increase was similarly suppressed in the group administered with the positive reference substance diltiazem (1 △ 1). The inhibitory effect of the diltiazem administration group was slightly stronger than that of the EMIQ administration group.

[0076] As shown in Fig. 4, the mean value of the diastolic blood pressure of the control group (-〇-) showed 94 mmHg before the start of administration, and then increased with aging, and on the 36th day after administration. Showed 163 mmHg. In the EMIQ administration group (1 mouth-), the value was lower than that in the control group after 8 days of administration, and the increase was suppressed. The increase was also similarly suppressed in the group administered with the positive control substance diltiazem (1 △ 1).

[0077] 2.3 Changes in heart rate

Figure 5 shows the heart rate measurement results. The average value of heart rate in the control group (-O-) was 426 beat / min before the start of administration, but thereafter decreased with aging, and showed 384 beat / min on the 36th day of administration. In the EMIQ administration group (one mouthful) and the diltiazem administration group (one △ one), a significant low value was observed on the 22nd day of administration, but thereafter there was a significant change compared to the control group. There wasn't.

[0078] 4. Discussion

This time, diltiazem used as a positive control substance is a calcium antagonist with antihypertensive action. Narita et al. (Hiroshi Narita et al .: Effects of Diltiazem on SHR blood pressure increase and associated cardiac hypertrophy and blood vessel thickening, Journal of Japanese Pharmacology 86 (3): 165-174, 1985) It was reported that the administration of diltiazem for 4 to 12 weeks after birth significantly suppressed the increase in blood pressure in SHR rats, but the heart rate was not different from the control group. In this study, although diltiazem showed antihypertensive activity, it did not affect the heart rate, and the same results as the above report were obtained.

[0079] The EMIQ used in the present invention is similar to diltiazem in terms of systolic blood pressure and mean blood pressure. In addition, each of the blood pressures of the diastolic blood pressure exhibited an action of suppressing the increase thereof. Furthermore, no adverse effects on heart rate, weight, etc. were observed by administration of EMIQ.

[0080] From these results, it was revealed that EMIQ has an effect of suppressing blood pressure increase in subjects who tend to develop hypertension or subjects who tend to develop hypertension. E MIQ's antihypertensive action was milder than dilutiazem, an antihypertensive agent (medicine), and it had no adverse effects on the body. It is considered to be particularly suitable for.

[0081] Test Example 2 Effects of EMIQ and quercetin on blood pressure

In the same manner as in Test Example 1, EMIQ prepared in Preparation Example 1 (2) and quercetin (QC), which is an aglycon of EMIQ (Preparation Example 1 (3)), were each applied to 5 weeks old spontaneously hypertensive rats. Oral administration was repeated twice a day for 7 weeks, and the inhibitory effect on blood pressure increase was examined mainly using changes in systolic blood pressure as an index (10 mice in each group). The dose of EMIQ is 26 mg / kg / day (13 mg / kg twice a day), and the dose of QC is 10.4 mg / kg / day (5.2 mg / kg is equivalent to 1 mol of EMIQ). Twice a day). As a control study, in parallel with these, no EMIQ and QC were administered, and systolic blood pressure was measured even in spontaneously hypertensive rats (non-administered group).

[0082] The results are shown in FIG. From this result, EMIQ is superior to QC in suppressing the increase in systolic blood pressure! /, Confirmed that.

[0083] Example 1 Food composition (1)

Using EMIQ prepared in Preparation Example 1 as an active ingredient having antihypertensive action, a beverage containing 20% drape juice was prepared. Specifically, the ingredients were mixed according to the following formulation, filtered, filled into a 250 ml bottle, sterilized at 80 ° C. for 10 minutes to prepare a beverage containing 20% grape juice. In the following examples, * indicates that the product is manufactured by San-Ei Gen F.F., Ltd., and * indicates that it is a registered trademark of San-Ei Gen F.F. [0084] Gu Prescription>

1.Sucralose * 0.0136 (kg)

2.5 times concentrated grape clear juice (Sanyo Foods Co., Ltd.) 4.00

3.Chenic acid (crystal) * 0.25

4. EMIQ 0.002

5.Grape flavor * 0.10

6. Funole flavor * 0.10

7.Water proper amount

Total 100.00 kg.

[0085] Jewelery 12 Food composition (2)

Using EMIQ prepared in Preparation Example 1 as an active ingredient having antihypertensive action, a spad link was prepared. Specifically, the materials were mixed according to the following formulation, filtered, and hot-packed into 500 mL PET bottles to prepare sports drinks.

[0086] Gu Prescription>

1.Sand sugar 4.000 (kg)

2.Fructose glucose liquid sugar 3.000

3.Chenic acid (anhydrous) medium grain * 0.200

4. Trisodium citrate fine granules * 0.085

5 丄 ascorbic acid (crystal) * 0.100

6.Food salt 0.065

7. Shiohi potassium (Tonda Pharmaceutical Co., Ltd.) 0.038

8. Calcium lactate (Taihei Chemical Industry Co., Ltd.) 0.015

9. Shiohig magnesium (Tonda Pharmaceutical Co., Ltd.) 0.005

10. Grapefruit flavor * 0.130

11. EMIQ 0.020

Total 100.000 L with water. Decor 13 Food composition (3)

A nutritional drink was prepared using the EMIQ prepared in Preparation Example 1 as an active ingredient having an antihypertensive action. Specifically, 1-3 were stirred and dissolved in water, and the total amount was corrected. Next, other ingredients were sequentially added at a temperature of 93 ° C., and hot packs were filled in lOOmL brown glass bottles to prepare energy drinks. <Prescription>

1. Fructose glucose liquid sugar 24.00

2.Chenic acid (anhydrous) * 0.55

3. Trisodium citrate * 0.55

4. Thiamine nitrate (Sankyo Lifetech Co., Ltd.) 0.005

5. Riboflavin (Daiichi Fine Chemical Co., Ltd.) 0.01

6. Pyridoxine hydrochloride (Daiichi Fine Chemical Co., Ltd.) 0.005

7.Lemon flavor * 0.20

8. EMIQ 0.30

Water total 100.00 L, Decorative 14 Food composition (4)

A non-sugar candy was prepared using the EMIQ prepared in Preparation Example 1 as an active ingredient having an antihypertensive action. Specifically, the materials were mixed according to the following formulation, heated and dissolved, and then molded to prepare a non-sugar candy.

[0090] <Prescription>

1. Powdered palatinit PNP (manufactured by Shin Mitsui Sugar Co., Ltd.) 60.00 (kg)

2. Amarty syrup (Towa Kasei Kogyo Co., Ltd.) 59.33

3.Sucralose * 0.03

4.Chenic acid (anhydrous) * 1.50

5. Trisodium citrate * 0.07

6. EMIQ 10.00

7. Grape flavored 0.20

8.Water ― _ 30.00

Boiled, total 100.00 kg.

[0091] Example 5 food composition (5)

A non-sugar candy was prepared using the EMIQ prepared in Preparation Example 1 as an active ingredient having an antihypertensive action. Specifically, the materials were mixed according to the following formulation, heated and dissolved, and then molded to prepare a non-sugar candy. <Prescription>

1. Powdered palatinit PNP (manufactured by Shin Mitsui Sugar Co., Ltd.) 70.00 (kg)

2. Amarty syrup (Towa Kasei Kogyo Co., Ltd.) 59.33

3. Sucralose rice cake 0.03

4.Chenic acid (anhydrous) * 1.50

5. Trisodium citrate * 0.07

6. EMIQ 0.1

7.Umeflano * 0.2

8.Water ― 30.0

Boiled, total 100.0 kg. Example 6 Food Composition (6)

A drink jelly was prepared using the EMIQ prepared in Preparation Example 1 as an active ingredient having an antihypertensive action. Specifically, the materials were mixed according to the following formulation, dissolved by heating, filled into containers, and sterilized at 85 ° C for 30 minutes to prepare drink jelly.

1.sugar 5.0 (kg)

2. Fructose glucose liquid sugar 15. 0

3. 5 times concentrated white grape juice (Sanyo Foods Co., Ltd.) 4.0

4.Gel-up * K-one S * 0.2

5. Gel-up * SA— 3C * 0. 3

6. Trisodium citrate * 0. 1

7. Calcium lactate (Showa Kako Co., Ltd.) 0.1

8.Chenic acid (anhydrous) * 0. 18

9. EMIQ 0.2

10. Grape flavor * 0.3

11.Water proper amount

100 kg.

Decoration 17 Food Composition (7)

A drink jelly was prepared using the EMIQ prepared in Preparation Example 1 as an active ingredient having an antihypertensive action. Specifically, the materials were mixed according to the following formulation, dissolved by heating, filled into containers, and sterilized at 85 ° C for 30 minutes to prepare drink jelly. [0096] Prescription>

1.sugar 5.0 (kg)

2. Fructose glucose liquid sugar 15. 0

3. 5 times concentrated white grape juice (Sanyo Foods Co., Ltd.) 4.0

4.Gel up * K-S * 0.2

5 Gel-up * SA— 3C * 0. 3

6. Trisodium citrate * 0. 1

7. Calcium lactate (Showa Kako Co., Ltd.) 0.1

8.Chenic acid (anhydrous) * 0. 18

9. EMIQ 0. 03

10. Grape flavor * 0.3

11.Water proper amount

π ρ Γ 100. 00 kg ₀ Example 8 Food composition (8)

Using the EMIQ prepared in Preparation Example 1 as an active ingredient having an antihypertensive action, materials were mixed according to the following formulation, and tablets were prepared using a tableting machine.

[0098] Gu Prescription>

1.Sorbit W—Powder (Towa Kasei Kogyo Co., Ltd.) 79.75 (kg)

2. DK Ester F-20W (Daiichi Kogyo Seiyaku Co., Ltd.) 0.15

3. Lemon flavor 氺 0.1

—4.—EMIQ 20.0

Total 100.00 kg. Decoration 19 Food composition (9)

[0100] <Prescription>

1.Sorbit W—Powder (Towa Kasei Kogyo Co., Ltd.) 95.75 (kg)

2. DK Ester F-20W (Daiichi Kogyo Seiyaku Co., Ltd.) 0.15

3. Yogurt flavor * 0.1

4. EMIQ 4.0

Total 100.0 kg. Example 10 Food composition (10)

Soft biscuits were prepared using the EMIQ prepared in Preparation Example 1 as an active ingredient having antihypertensive action. Specifically, 3 and 6 are mixed in a universal mixing stirrer using a beater with 126 r pmZl for 30 seconds, then 2 and 5 are added, and further mixed at 126 rpm for 3 minutes. Then 12 While mixing at 6 rpm, add 4 little by little, then mix 1 and 7 that have been sieved. After laying in a refrigerator for 30 minutes, roll to 15mm thickness, cut to 20mm x 30mm, and mold. A soft biscuit was prepared by baking for about 30 minutes in an oven at 150 ° C and 120 ° C.

Prescription>

1. Soft flour

2. White sugar 35.0

3.Margarine 40.0

4. Whole egg 20.0

5. Nonfat dry milk 10.0

6. Toasted butter oil NO.46970 * 0.2

7. EMIQ 0.3

Total (before firing) 205.5 kg.

Brief Description of Drawings

o

[Fig. 1] Each sample was administered to each group [control group (10-), enzyme-treated isoquercitrin (EMIQ) -administered group (mouthful--), and diltiazem-administered group (positive control group) (1- △-)]. FIG. 3 is a graph showing changes in body weight over time (Test Example 1).

[Fig. 2] In Test Example 1, each group [control group (10—), enzyme-treated isoquercitrin (EMIQ) administration group (one mouthful), and diltiazem administration group (positive control group) (one △ one)] Is a diagram showing the results of systolic blood pressure measured after administration of each sample. In the figure, ** indicates a significant difference from the control group of 0.01 and # indicates that the significant difference from the EMIQ administration group is P of 0.05 (same in FIGS. 3 to 5).

[Fig. 3] In Test Example 1, each group [control group (100-), enzyme-treated isoquercitrin (EMIQ) administration group (one mouthful), and diltiazem administration group (positive control group) (one △ one)] Is a diagram showing an average value of average blood pressure measured after administration of each sample.

[Fig. 4] In Test Example 1, each group [control group (10—), enzyme-treated isoquercitrin (EMIQ) administration group (one mouthful), and diltiazem administration group (positive control group) (one △ one)] Is a diagram showing the results of diastolic blood pressure measured after administration of each sample. In the figure, * indicates that the significant difference from the control group is P 0.05 (the same applies to FIG. 5).

[Fig. 5] In Test Example 1, each group [control group (10—), enzyme-treated isoquercitrin (EMIQ ) In the administration group (one mouthful) and the diltiazem administration group (positive control group) (one Δ1)], it is a graph showing the heart rate measured after administration of each sample.

FIG. 6 shows the results of measuring changes in systolic blood pressure over time for the enzyme-treated isoquercitrin (EMIQ) administration group, quercetin (QC) administration group, and non-administration group (control) in Test Example 2. FIG. In the figure, ** indicates that the significant difference from the control group is 0.01, and $$ indicates that the significant difference from the E MIQ administration group is 0.01.

Claims

The scope of the claims

[I] An antihypertensive composition comprising enzyme-treated isoquercitrin as an active ingredient.

[2] The enzyme-treated isoquercitrin contains an effective amount that exhibits an antihypertensive effect.

1. The antihypertensive composition according to 1.

[3] The antihypertensive composition according to claim 1, having an oral dosage form.

[4] The antihypertensive composition according to claim 1, containing enzyme-treated isoquercitrin at a ratio of 1 mg to LOg per daily dosage unit.

[5] The antihypertensive composition according to claim 1, wherein the enzyme-treated isoquercitrin is formulated so as to contain 1 mg to LOg per daily dosage unit.

6. The antihypertensive composition according to claim 1, which is a pharmaceutical product.

7. The antihypertensive composition according to claim 1, which is a food.

[8] The antihypertensive composition according to claim 7, wherein the antihypertensive composition is contained in a container, and the antihypertensive effect is described as an effect on the container or the package thereof.

[9] The food for specified health use for preventing the onset of hypertension or suppressing the development of hypertension

7. The antihypertensive composition described in 7.

[10] A method for preventing the onset of hypertension or suppressing the development of hypertension, comprising administering to a subject at risk of hypertension an effective amount of enzyme-treated isoquercitrin that exhibits an antihypertensive effect.

[II] Use of enzyme-treated isoquercitrin for the production of an antihypertensive composition.