JP2023033087A - Vascular flexibility improving composition, vascular endothelial function improving composition, platelet aggregation inhibitory composition, blood flow improving composition, and oral composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide compositions having excellent vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action, and useful as an oral composition.

SOLUTION: A composition contains (A) catechin and (B) procyanidin B3. In the composition, the ratio of the content of the component (A) to the content of the component (B) [(A)/(B)] is 0.60 or more and 15 or less.

SELECTED DRAWING: Figure 1

COPYRIGHT: (C)2023,JPO&INPIT

Description

The present invention provides a composition containing (A) catechin and (B) procyanidin B3 (hereinafter also referred to as "PB3"), wherein the content of component (B) in the composition A composition for improving vascular flexibility, a composition for improving vascular endothelial function, and a platelet aggregation inhibitor, wherein the content ratio of (A) [(A)/(B)] is 0.60 or more and 15 or less It relates to a composition for blood flow, a composition for improving blood flow, and an oral composition.

In recent years, lifestyle-related diseases are increasing with changes in eating habits and lifestyles. Lifestyle-related diseases include diseases such as hypertension, hyperlipidemia, and diabetes, as well as diseases such as angina pectoris, myocardial infarction, and cerebral circulation disorders, whose onset can be prevented by improving lifestyle habits. is a generic term for As one of the factors of these diseases, it has been clarified that decreased vascular flexibility, decreased vascular endothelial function, thrombus formation due to increased platelet aggregation, and decreased blood flow are involved. Moreover, since lifestyle-related diseases are chronic diseases, it is necessary to require a long period of time for their prevention and treatment. Therefore, it is expected to develop a food that can safely and effectively improve vascular flexibility, vascular endothelial function, inhibit platelet aggregation, and improve blood flow over a long period of time. For example, hops (Patent Document 1), chlorogenic acid (Patent Document 2), and piperine (Patent Document 3) are known as food materials having such effects.

JP 2005-104951 JP 2003-261444 JP 2020-090551

However, according to investigations by the present inventors, hops, chlorogenic acid, and piperine do not have sufficient vascular flexibility-improving effects, vascular endothelial function-improving effects, platelet aggregation-inhibiting effects, and blood flow-improving effects.

Accordingly, the present inventors focused on (A) catechin and (B) procyanidin B3, and conducted various studies with the aim of effectively utilizing these components.

As a result of intensive investigation and research, the present inventors have found that by adjusting the content of (A) catechin and (B) procyanidin B3 to a specific ratio, excellent vascular flexibility improving action and vascular endothelial function improvement , platelet aggregation inhibitory action, and blood flow improving action. The invention is a completed invention based on such findings.

The outline of the present invention is as follows.
[1] A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for improving vascular flexibility, wherein the content of component (A) relative to the content of component (B) in the composition A composition characterized in that the content ratio of [(A)/(B)] is 0.60 or more and 15 or less.
[2] A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for improving vascular endothelial function, wherein the content of component (A) relative to the content of component (B) in the composition A composition characterized in that the content ratio of [(A)/(B)] is 0.60 or more and 15 or less.
[3] A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for inhibiting platelet aggregation, wherein the ratio of component (A) to the content of component (B) in the composition A composition characterized in that the content ratio [(A)/(B)] is 0.60 or more and 15 or less.
[4] A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for improving blood flow, wherein the ratio of component (A) to the content of component (B) in the composition A composition characterized in that the content ratio [(A)/(B)] is 0.60 or more and 15 or less.
[5] An oral composition containing (A) catechin and (B) procyanidin B3, wherein the ratio of the content of component (A) to the content of component (B) in the composition [(A)/ (B)] is 0.75 or more and 10.3 or less.
[6] A composition for improving vascular flexibility containing (A) catechin and (B) procyanidin B3, wherein the ratio of the content of component (A) to the content of component (B) in the composition [ (A)/(B)] is 0.60 or more and 15 or less.
[7] A composition for improving vascular endothelial function containing (A) catechin and (B) procyanidin B3, wherein the ratio of the content of component (A) to the content of component (B) in the composition [ A composition for improving vascular endothelial function, wherein (A)/(B)] is 0.60 or more and 15 or less.
[8] A composition for inhibiting platelet aggregation containing (A) catechin and (B) procyanidin B3, wherein the ratio of the content of component (A) to the content of component (B) in the composition [( A)/(B)] is 0.60 or more and 15 or less.
[9] A composition for improving blood flow containing (A) catechin and (B) procyanidin B3, wherein the ratio of the content of component (A) to the content of component (B) in the composition [( A)/(B)] is 0.60 or more and 15 or less.
[10] The ratio [(A)/(B)] of the content of component (A) to the content of component (B) in the composition is 0.75 or more and 10.3 or less [ 1] to the composition according to any one of [9].
[11] The composition according to any one of [1] to [10], which contains a pine bark extract.

According to the present invention, there is provided a composition containing (A) catechin and (B) procyanidin B3. It is possible to provide a composition useful as an oral composition, which is excellent in flexibility-improving action, vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action.

FIG. 1 shows the expression levels of NO synthase genes in Examples and Comparative Examples. FIG. 2 shows the amount of prostaglandin I2 produced in Examples and Comparative Examples.

The composition for improving vascular flexibility, the composition for improving vascular endothelial function, the composition for inhibiting platelet aggregation, the composition for improving blood flow, and the oral composition of the present invention (hereinafter collectively referred to as "the composition of the present invention"). (also referred to as "composition") contains (A) catechin and (B) procyanidin B3, and is characterized by a specific ratio between the content of (A) catechin and the content of (B) procyanidin B3. The composition of the present invention may further contain other ingredients, if desired. Each component and the composition of the present invention are described below. It should be noted that the configuration described below does not limit the present invention, and various modifications can be made within the scope of the gist of the present invention.

The composition of the present invention contains (A) catechin. Catechin is a compound represented by the chemical formula C15H14O6, and the catechin in the present invention is (+)-catechin represented by the following chemical formula (1) or (-)-catechin represented by the chemical formula (2). As the catechin, either (+)-catechin or (-)-catechin may be used, or a combination of the two may be used. From the viewpoint of inhibitory action and blood flow improving action, it is preferable to use the two in combination.

The catechin of the present invention can be a synthetic product or a plant-derived product such as pine, tea, etc. From the viewpoint of safety during use, plant-derived products are preferable, and pine is more preferable because it contains abundant catechins. Particular preference is given to using pine bark.

The blending ratio of catechins blended in the composition of the present invention is not particularly limited, and the blending ratio can be appropriately set in a wide range according to various conditions such as the purpose, shape, and intended use. For example, when the composition of the present invention is a solid agent (powder, granule, granule, tablet, capsule, chewable, etc.), it is preferably 0.000001 to 5.0% by mass, and blood vessel flexibility. 0.00001 to 3.0% by mass is more preferable, and 0.0001 to 1.0% by mass is particularly preferable, from the viewpoint of improving action, vascular endothelial function improving action, platelet aggregation inhibiting action, and blood flow improving action. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 0.000001 to 1.0% by mass, and 0.00001 to 0.0001 0.1 mass % is more preferred, and 0.0001 to 0.01 mass % is particularly preferred.

The amount of catechins in the composition of the present invention can be analyzed by HPLC. For example, it can be measured by an HPLC analyzer equipped with an ultraviolet absorption detector, the analysis column is L-Column ODS 3 μm (4.6 × 250 mm) manufactured by the Chemicals Evaluation and Research Institute, and the mobile phase liquid As media, 0.1 M acetic acid aqueous solution (mobile phase A) and 0.1 M acetonitrile acetate solution (mobile phase B) can be used, the column temperature can be 40° C., and the flow rate can be 1.0 mL/min. Gradient conditions can be as follows.

Pine that can be used as a source of catechin in the present invention includes, for example, French maritime pine, larch, black pine, Japanese red pine, Japanese white pine, Japanese pine, Korean pine, creeping pine, Ryukyu pine, Japanese pines, Giant pine, White pine, Aneda in the Quebec region of Canada, and the like. plants belonging to the order Pinaceae, but are not limited thereto. Among these, French maritime pine (Pinus pinaster), which is a marine pine growing on the Atlantic coast of southern France, etc., is preferable because it has been confirmed to be safe for food and contains a high concentration of catechins. .

The method of processing the plant-derived material is not particularly limited, and processed materials such as pulverized materials, squeezed juices, and extracts can be used. Powders, granules and the like can be mentioned as pulverized products. The squeezed juice or extract may be in liquid form, but may also be used in the form of paste or dry powder. When a paste or dry powder is used, it may be produced by using only the substance itself, or may be processed together with an excipient. In the present invention, it is preferably obtained by extraction from the viewpoint of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibiting action, and blood flow improving action. Hereinafter, a method for extracting a plant-derived substance will be described using a method for producing a pine bark extract as an example.

A pine bark extract is obtained by extracting pine bark with a solvent. Examples of the solvent include, but are not limited to, water, organic solvents, and water-containing organic solvents (water-containing alcohols such as water-containing ethanol). As the organic solvent used for extraction, an organic solvent that is generally acceptable for extracting natural product components is used. Acetone, hexane, cyclohexane, propylene glycol, hydrous ethanol, hydrous propylene glycol, ethyl methyl ketone, glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible oil, 1,1,1,2-tetrafluoroethane, 1, 1,2-trichloroethene and the like. These solvents may be used singly or in combination of two or more. The temperature during extraction can be appropriately adjusted from room temperature to a temperature below the boiling point of the extraction solvent. In the present invention, water, hydrous ethanol, and hydrous propylene glycol are preferably used from the viewpoint of efficiently extracting catechins.

The extraction method is not particularly limited as long as it is a method generally accepted for extracting components of natural products, and examples thereof include solid-liquid extraction methods such as hot extraction and supercritical fluid extraction.

The heated extraction method is, for example, a method of contacting a test substance with a solvent, treating at a temperature below the boiling point of the solvent, and extracting components contained in the test substance into the solvent. A reflux extraction method may also be used.

The supercritical fluid extraction method is, for example, a method of performing extraction using a supercritical fluid, which is a fluid in a state exceeding the critical point (critical temperature, critical pressure) of the gas-liquid of the substance. Examples of supercritical fluids include carbon dioxide, ethylene, propane, nitrous oxide (laughing gas), etc. Carbon dioxide is preferred.

In the supercritical fluid extraction method, an extraction step of extracting a target component with a supercritical fluid and a separation step of separating the target component and the supercritical fluid are performed. In the separation step, any of extraction and separation by pressure change, extraction and separation by temperature change, and extraction and separation using an adsorbent/absorbent may be performed.

Supercritical fluid extraction by the entrainer addition method may be performed. This method uses, for example, ethanol, propanol, n-hexane, acetone, toluene, other aliphatic lower alcohols, aliphatic hydrocarbons, aromatic hydrocarbons, and ketones in the extraction fluid at 2 to 20 W/V. % and performing supercritical fluid extraction using this fluid, the solubility of the desired extract such as OPC (oligomeric proanthocyanidin) and catechins in the extraction solvent is dramatically increased. or to enhance the selectivity of separation, and to obtain an efficient pine bark extract.

Supercritical fluid extraction can be operated at relatively low temperatures, so it has the advantage of being applicable to substances that degrade or decompose at high temperatures. There is an advantage that the process can be omitted and the process becomes simple.

Extraction from pine bark may be performed by a liquid carbon dioxide batch method, a liquid carbon dioxide reflux method, a supercritical carbon dioxide reflux method, or the like, in addition to the extraction method described above.

Extraction from pine bark may combine multiple extraction methods. By combining multiple extraction methods, it is possible to obtain pine bark extracts with various compositions.

It is safe to purify the pine bark extract obtained by extraction by ultrafiltration, a column method using an adsorptive carrier (Diaion HP-20, Sephadex-LH20, chitin, etc.), a batch method, or the like. preferable from the aspect.

The pine bark extract is preferably in the form of powder, more preferably in the form of dry powder, from the viewpoint of storage stability and processability. The drying means is not particularly limited, and examples thereof include drying the solvent-containing pine bark extract by heating, sun-drying, hot-air drying, freeze-drying, reduced pressure, and the like. The degree of drying may be until it is confirmed that the solvent content of the pine bark extract is sufficiently reduced, for example, until the solvent content is 10 wt% or less, preferably 5 wt% or less. degree.

Examples of the powdering method for obtaining the pine bark extract dry powder include a method of pulverizing and pulverizing the pine bark extract with a ball mill, hammer mill, roller mill, etc., which are methods commonly used by those skilled in the art. include but are not limited to: It is also possible to change the order of drying and pulverization, pulverize the pine bark extract before drying, and dry the pulverized product to obtain a pine bark extract dry powder.

Commercially available pine bark extracts may be used, and examples of commercially available pine bark extracts include flavangenol (registered trademark; Toyo Shinyaku Co., Ltd.).

The composition of the present invention contains (B) procyanidin B3 (PB3). Procyanidin B3 is a C4-C8-linked dimer of (+)-catechin and (+)-catechin. PB3) of the present invention can be a synthetic product or a plant-derived product such as pine, apple, cocoa or grape, and from the viewpoint of safety during use, a plant-derived product is preferable, pine is more preferable, and PB3 is abundant. It is particularly preferred to use pine bark because it contains For the types of pine and the processing method of the plant-derived material that can be used in the composition of the present invention, reference can be made to the content described in Catechin.

The blending ratio of PB3 blended in the composition of the present invention is not particularly limited, and the blending ratio can be appropriately set in a wide range according to various conditions such as the purpose, shape, and intended use. For example, when the composition of the present invention is a solid agent (powder, granule, granule, tablet, capsule, chewable, etc.), it is preferably 0.000001 to 5.0% by mass, and blood vessel flexibility. 0.00001 to 3.0% by mass is more preferable, and 0.0001 to 1.0% by mass is particularly preferable, from the viewpoint of improving action, vascular endothelial function improving action, platelet aggregation inhibiting action, and blood flow improving action. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 0.000001 to 1.0% by mass, and 0.00001 to 0.0001 0.1 mass % is more preferred, and 0.0001 to 0.01 mass % is particularly preferred.

The amount of PB3 in the composition of the invention can be analyzed by HPLC. For example, it can be measured by an HPLC analyzer equipped with an ultraviolet absorption detector, the analysis column is L-Column ODS 3 μm (4.6 × 250 mm) manufactured by the Chemicals Evaluation and Research Institute, and the mobile phase liquid As media, 0.1 M acetic acid aqueous solution (mobile phase A) and 0.1 M acetonitrile acetate solution (mobile phase B) can be used, the column temperature can be 40° C., and the flow rate can be 1.0 mL/min. Gradient conditions can be as follows.

The composition of the present invention contains (A) catechin and (B) procyanidin B3, and is characterized in that the content of (A) catechin and the content of (B) procyanidin B3 are in a specific ratio.

The composition of the present invention contains (A) catechin and (B) procyanidin B3, and the ratio of the content of component (A) to the content of component (B) in the composition [(A)/(B) ] is 0.60 or more and 15 or less, it has excellent vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibiting action, and blood flow improving action. The lower limit of the ratio [(A)/(B)] of the content of component (A) to the content of component (B) is not limited as long as it is 0.60 or more, but is preferably 0.70 or more. , vascular flexibility-improving action, vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action, it is particularly preferably 0.75 or more. The upper limit of the ratio [(A)/(B)] of the content of component (A) to the content of component (B) is not limited as long as it is 15 or less, but is preferably 12 or less. It is particularly preferably 10.3 or less from the viewpoint of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibiting action, and blood flow improving action.

The ratio of the total amount of (A) catechin and (B) PB3 to be blended in the composition of the present invention is not particularly limited, and it can be used in a wide range depending on various conditions such as purpose, shape, and intended use. The mixing ratio can be appropriately set. For example, when the composition of the present invention is a solid formulation (powder, granule, granule, tablet, capsule, chewable, etc.), the content is preferably 0.000002 to 10% by mass, and the effect of improving blood vessel flexibility. , 0.00002 to 6.0% by mass is more preferable, and 0.0002 to 2.0% by mass is particularly preferable, from the viewpoint of vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 0.000002 to 2.0% by mass, and 0.00002 to 0.0 0.2 mass % is more preferred, and 0.0002 to 0.02 mass % is particularly preferred.

The daily intake per body weight of the composition of the present invention is not particularly limited, and can be appropriately set according to the mode of use, the details of use by the user, and the like. For example, when the composition of the present invention is a solid formulation (powder, granule, granule, tablet, capsule, chewable, etc.), it is preferably 0.2 to 6000 mg based on the weight of the user. /kg, more preferably 1 to 4000 mg/kg, and particularly preferably 2 to 2000 mg/kg in terms of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action. is. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 200 to 20000 mg/kg based on the body weight of the user. , more preferably 600 to 15,000 mg/kg, and particularly preferably 1,000 to 10,000 mg/kg in terms of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action.

Similarly, there is no particular limitation on the amount of the composition of the present invention that is ingested per body weight. For example, when the composition of the present invention is a solid formulation (powder, granule, granule, tablet, capsule, chewable, etc.), it is preferably 0.06 to 6000 mg based on the weight of the user. /kg, more preferably 0.3 to 4000 mg/kg, particularly preferably 0.6 from the viewpoint of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action ~2000 mg/kg. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 66 to 20000 mg/kg based on the body weight of the user. , more preferably 200 to 15,000 mg/kg, and particularly preferably 333 to 10,000 mg/kg in terms of vascular flexibility-improving action, vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action.

The daily intake of the composition of the present invention is not particularly limited. is preferably 0.01 to 300 g, more preferably 0.05 to 200 g, and is particularly preferably 0.0 g from the viewpoint of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action. It can be from 1 to 100 g. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 10 to 1000 g, more preferably 30 to 750 g. 50 to 500 g is particularly preferable from the viewpoint of action, vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action.

The amount of the composition of the present invention to be taken at one time is not particularly limited. is preferably 0.003 to 300 g, more preferably 0.01 to 200 g, particularly preferably 0.00 g from the viewpoint of vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action. 03-100 g. Further, for example, when the composition of the present invention is a liquid (liquid, gel, paste, jelly, yoghurt, etc.), it is preferably 3.3 to 1000 g, more preferably 10 to 750 g, blood vessel softening 16 to 500 g is particularly preferable from the viewpoint of sex improving action, vascular endothelial function improving action, platelet aggregation inhibiting action, and blood flow improving action.

The composition of the present invention may be taken before meals, between meals, after meals, or at the same time as meals. , preferably before meals, after meals, or with meals, more preferably before meals or with meals.

The composition of the present invention may contain only catechin and PB3, or may contain other ingredients in addition to these. Examples of the other ingredients include protein, dietary fiber such as water-soluble dietary fiber and insoluble dietary fiber, vitamins, minerals, algae, lactic acid bacteria, microorganisms such as yeast, and the like. Furthermore, if necessary, sugars such as dextrin and starch, oligosaccharides, sweeteners, acidulants, coloring agents, thickeners, brighteners, excipients, nutritional supplements, binders, and lubricants that are usually used in the food field. Lubricants, stabilizers, diluents, bulking agents, emulsifiers, food additives, seasonings and the like can be mentioned. The content of these other components can be appropriately selected according to the form of the composition of the present invention.

The composition of the present invention is particularly limited as long as it can be distinguished from other products as a product in that it can be used for improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, or improving blood flow. However, for example, the main body, packaging, instructions, or advertising materials of the product according to the present invention have a function of improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, or improving blood flow. Any indication to that effect is included within the scope of the present invention. Examples include pharmaceuticals (including quasi-drugs), food for specified health uses, food with nutrient function claims, food with functional claims, and other so-called health foods such as functional foods whose efficacy has been approved by a designated organization. can be done. In so-called health foods, it is useful to maintain the flexibility of blood vessels (degree of expansion of blood vessels after constriction), which declines with age, to improve blood vessel function, to maintain blood vessel function, and to maintain blood vessel function. Enhances endothelial function", "Maintains vascular endothelial function", "Enhances vascular endothelial function of arteries that decline with age", "Maintains vascular endothelial function of arteries that declines with age", "After constricting blood vessels" "Increase dilatation of blood vessels", "Maintain dilatation of blood vessels after tightening", "Enhance blood vessel flexibility", "Maintain blood vessel flexibility", "Increase flexibility of blood vessels" , "Maintains suppleness of blood vessels", "Maintains platelet aggregation ability", "Helps maintain blood smoothness (platelet aggregation ability)", "Helps maintain blood fluidity (platelet aggregation ability)" ", "helps maintain easy blood flow (platelet aggregation)", "improves blood flow", "maintains blood flow", "maintains healthy blood flow (peripheral blood flow)", " For example, it is possible to display the words such as "Restore decreased blood flow to normal by increasing peripheral blood flow" and "Restore decreased blood flow (peripheral blood flow) to normal".

The form of the composition of the present invention is not particularly limited, and can be any form. For example, forms suitable for oral use, specifically powders, granules, granules, tablets, liquids, gels, pastes, capsules such as hard capsules and soft capsules, caplets, tablets, Examples include gel, jelly, gummy, wafer, biscuit, cookie, cake, chewable, syrup, stick, and yoghurt forms. The composition of the present invention is preferably in the form of granules, tablets, capsules, or liquid from the viewpoint of vascular flexibility-improving action, vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action. A shape or a capsule shape is more preferable. Here, the term "granular" refers to a composition obtained by granulating powder, which may be drunk directly or dissolved in a liquid such as water. Moreover, the term "gel-like" refers to a composition containing water and a thickening agent or a gelling agent and having viscosity or elasticity.

The packaging form of the composition of the present invention is not particularly limited, and can be appropriately selected according to the dosage form. Examples include packaging, glass containers such as vials, and plastic containers such as ampoules.

The present invention will be described in more detail below with reference to Examples, but the present invention is not limited to these Examples, and the present invention can take various aspects as long as the problems of the present invention can be solved. .

<Test 1 Evaluation of NO synthase gene expression level>
In this test, the expression level of nitric oxide (NO) synthase gene was evaluated. When NO is produced by NO synthase (eNOS) in the vascular endothelium and diffused to reach the vascular smooth muscle, the muscle relaxes and the blood vessel dilates. When the expression level of the NO synthase gene increases, the amount of NO synthase increases and the amount of NO production increases. Therefore, improvement of vascular flexibility, improvement of vascular endothelial function, and improvement of blood flow can be expected. As described above, by evaluating the NO synthase gene expression level, it is possible to evaluate the effect of improving vascular flexibility, the effect of improving vascular endothelial function, and the effect of improving blood flow.

(1) Test substance As a test substance, (+)-catechin and (-)-catechin, pine bark extract containing PB3, (-)-catechin reagent (manufactured by Nagara Science), PB3 reagent (manufactured by ChemFaces) used. These are mixed, dissolved in DMSO so as to have the ratio shown in Table 1, and diluted with Endothelial Cell Growth Medium 2 (EGM-2) medium (manufactured by Lonza) so that the added concentration of DMSO is 0.5%. Thus, the test substances of Examples and Comparative Examples were prepared. (A) catechin is the total amount of (+)-catechin and (−)-catechin.

(2) Cell culture Human umbilical vein endothelial cells (manufactured by Lonza) were cultured in EGM-2 medium. The cells were seeded at 100 μL/well in a collagen-coated 96-well plate at 1×10 ⁴ cells/well, and precultured at 37° C. in a 5 vol % CO ₂ incubator for 24 hours. After removing the medium from each well, 100 μL/well of the test substance-containing medium shown in Table 1 was added and cultured for 24 hours.

(3) Measurement of NO synthase gene expression level After removing the medium from each well, the wells were washed once with PBS, and RNA was recovered using RNeasy Mini Kit (manufactured by QIAGEN). Real-time PCR was performed on the obtained RNA using One Step TBGreen (registered trademark) PrimeScript RT-PCR Kit II (manufactured by Takara). The NOS3 (eNOS) gene expression level was measured using a NOS3 (eNOS) primer (manufactured by Takara). As an endogenous control, GAPDH gene expression levels were measured using GAPDH primers (manufactured by QIAGEN).

(4) Evaluation of NO synthase gene expression level (B) The ratio of the content of (A) catechin to the content of PB3 [(A)/(B)] was 20.0 in the group (Comparative Example 2) A relative value of the NO synthase gene expression level when the expression level was set to 1 was calculated. The results are shown in FIG.

(B) The ratio of the content of (A) catechin to the content of PB3 (B) The group in which the ratio of the content of (A) catechin to the content of PB3 [(A)/(B)] is 0.75 (Example 1) and (B) the ratio of the content of (A) catechin to the content of PB3 (B) The ratio of the content of (A) catechin to the content of PB3 [(A) / (B)] In the group (Example 2) of 10.3, the group (Comparative Example 1 ) and (B) the ratio of the content of (A) catechin to the content of PB3 [(A) / (B)] compared to the group (Comparative Example 2) where the NO synthase gene expression level was 20.0 increased. Therefore, from this, the composition of the present invention is a composition in which the ratio [(A)/(B)] of the content of (A) catechin to the content of (B) PB3 is 0.60 or more and 15 or less. By ingestion, the effect of increasing the NO synthase gene expression level is exhibited, thereby exhibiting vascular flexibility improving action, vascular endothelial function improving action, platelet aggregation inhibitory action, and blood flow improving action. It was suggested that it is useful as

<Test 2 Evaluation of prostaglandin I2 production>
In this test, prostaglandin I2 (PGI2) production was evaluated. When prostaglandin I2 is produced in the vascular endothelium and diffuses to reach the vascular smooth muscle, it relaxes the muscle and dilates the blood vessel. An increase in prostaglandin I2 production is expected to improve vascular flexibility, vascular endothelial function, inhibit platelet aggregation, and improve blood flow. As described above, by evaluating the amount of prostaglandin I2 produced, it is possible to evaluate the effect of improving vascular flexibility, the effect of improving vascular endothelial function, the effect of inhibiting platelet aggregation, and the effect of improving blood flow. Since prostaglandin I2 has a short half-life, 6-keto prostaglandin F1α, which is a metabolite of prostaglandin I2, was evaluated for prostaglandin I2 production.

(1) Test substance A test substance was prepared in the same manner as in “Test 1 (1) Test substance”.

(2) Cell culture Cells were cultured in the same manner as in "Test 1 (2) Cell culture".

(3) Measurement of Prostaglandin I2 Production Medium was collected from each well, and the amount of 6-keto Prostaglandin F1α in the medium was measured using a 6-keto Prostaglandin F1α ELISA kit (manufactured by Cayman).

(4) Evaluation of prostaglandin I2 production (B) The ratio of the content of (A) catechin to the content of PB3 [(A)/(B)] of the group (Comparative Example 2) of 20.0 The relative value of prostaglandin I2 production was calculated when the expression level was set to 1. The results are shown in FIG.

(B) Group (Example 1) in which the ratio of (A) catechin content to PB3 content [(A)/(B)] is 0.75 and (B) (A) to PB3 content In the group (Example 2) having a catechin content ratio [(A)/(B)] of 10.3, the catechin content ratio [(A) /(B)] is 0.50 (Comparative Example 1) and the ratio of the content of (A) catechin to the content of (B) PB3 [(A)/(B)] is 20.0 An increase in prostaglandin I2 production was observed compared to the group (Comparative Example 2). Therefore, from this, the composition of the present invention is a composition in which the ratio [(A)/(B)] of the content of (A) catechin to the content of (B) PB3 is 0.60 or more and 15 or less. By ingestion, the effect of increasing the amount of prostaglandin I2 production is exhibited, thereby exhibiting an action of improving vascular flexibility, an action of improving vascular endothelial function, an action of inhibiting platelet aggregation, and an action of improving blood flow. It was suggested that it is useful as

(Manufacturing example)
Based on the results of Examples, Production Examples of the present invention are shown below.

[Production Example 1-3: Granules 1]
Granules 1 containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 were produced according to the formulation shown in Table 2. The granules described in Production Example 1-3 may be ingested in an amount of 3 g per day, and may be ingested after being dissolved in a solvent such as 100 ml of water, or may be ingested without being dissolved. All of the granules of Production Examples 1-3 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 4-6: Granules 2]
Granules 2 containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 were produced according to the formulation shown in Table 3. The granules described in Production Examples 4-6 should be taken in an amount of 25 g per day, and can be taken by dissolving in 200 ml of water or the like. All of the granules of Production Examples 4-6 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 7-9: Tablet]
Tablets containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 were manufactured according to the formulation in Table 4. Tablets were manufactured to have a tablet diameter of 8 mmφ, a tablet thickness of 4.5 mm, a weight of 250 mg, and a hardness of 5 kgf or more. One or two tablets described in Production Examples 7-9 may be ingested per day, and can be ingested with 100 ml of water or the like. All of the tablets of Production Examples 7-9 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 10-12: Hard Capsule]
By encapsulating the contents of the formulation of Table 5 with a coating containing gelatin or hydroxypropyl cellulose, (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 are contained. A hard capsule was produced. A hard capsule was manufactured with 300 mg per tablet. One or two tablets may be taken per day, and can be taken with 100 ml of water or the like. In addition, all of the hard capsules of Production Examples 10-12 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 13-15: Soft capsule]
Soft capsules containing (A) catechin (containing (+)-catechin and (−)-catechin) and (B) PB3 were produced by encapsulating the contents of the formulation in Table 6 with a gelatin-containing coating. bottom. Soft capsules were manufactured with 300 mg per tablet. One or two tablets may be taken per day, and can be taken with 100 ml of water or the like. In addition, all of the soft capsules of Production Examples 13-15 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Examples 16-18: PET beverage]
A PET beverage containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 was produced according to the formulation in Table 7. PET beverages were produced in 500 ml bottles. One should be taken per day. In addition, all of the PET beverages of Production Examples 16-18 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 19-21: Chocolate]
A chocolate containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 was produced according to the formulation in Table 8. You should take 30g per day. In addition, any of the chocolates of Production Examples 19 to 21 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Examples 22-24: Jelly]
A jelly containing (A) catechin (containing (+)-catechin and (−)-catechin) and (B) PB3 was produced according to the composition shown in Table 9. You should take 150g per day. In addition, all of the jelly of Production Examples 22-24 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 25-27: Gummy]
Gummies containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 were produced according to the formulation in Table 10. You should take 4g per day. In addition, all of the gummies of Production Examples 25-27 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

[Production Example 28-30: Yogurt]
A yogurt containing (A) catechin (containing (+)-catechin and (-)-catechin) and (B) PB3 was produced according to the formulation in Table 11. You should take 150g per day. In addition, all of the yogurts of Production Examples 28-30 are effective in improving vascular flexibility, improving vascular endothelial function, inhibiting platelet aggregation, and improving blood flow, and are useful as oral compositions.

INDUSTRIAL APPLICABILITY The composition of the present invention exhibits excellent vascular flexibility-improving action, vascular endothelial function-improving action, platelet aggregation-inhibiting action, and blood flow-improving action, and is therefore useful as an oral composition and industrially useful. is expensive.

Claims (5)

A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for improving vascular flexibility, wherein the content of component (A) relative to the content of component (B) in the composition A composition characterized in that the ratio of [(A)/(B)] is 0.60 or more and 15 or less. A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for improving vascular endothelial function, wherein the content of component (A) relative to the content of component (B) in the composition A composition characterized in that the ratio of [(A)/(B)] is 0.60 or more and 15 or less. A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for inhibiting platelet aggregation, wherein the content of component (A) relative to the content of component (B) in the composition A composition characterized in that the ratio [(A)/(B)] is 0.60 or more and 15 or less. A composition containing (A) catechin and (B) procyanidin B3 as an active ingredient for improving blood flow, wherein the content of component (A) relative to the content of component (B) in the composition A composition characterized in that the ratio [(A)/(B)] is 0.60 or more and 15 or less. (A) An oral composition containing catechin and (B) procyanidin B3, wherein the ratio of the content of component (A) to the content of component (B) in the composition [(A)/(B) ] is 0.60 or more and 15 or less.

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