Composition for promoting nitric oxide production

Abstract

The present invention relates to a composition for promoting nitric oxide production. [Problem] To provide a composition for promoting nitric oxide production. [Solution] Provided is a composition for promoting nitric oxide production, comprising rhizoma polygonati and / or fennel.

Description

Technical Field

[0001] The present invention relates to a composition for promoting the production of nitric oxide, which comprises polygonatum sibiricum and / or fennel. Background Art

[0002] It is known that vascular endothelial cells are cells that form the inner surface of blood vessels and have the function of producing bioactive substances such as nitric oxide (NO). Nitric oxide is one of the relaxation factors derived from vascular endothelium and is produced in vivo as a by-product of the conversion reaction in which L-arginine is converted to L-citrulline using nitric oxide synthase (NOS). Nitric oxide is a molecule that contributes to vascular function, for example, it can improve the elasticity of blood vessels and is less likely to cause arteriosclerosis. Therefore, promoting the production of nitric oxide in vascular endothelial cells is effective for relaxing and dilating blood vessels and inhibiting arteriosclerosis.

[0003] As a method for increasing nitric oxide, drugs such as statins and angiotensin II receptor antagonists (ARBs) are known, but it is effective for daily prevention of arteriosclerosis and the like to take them in the form of food on a daily basis. So far, isoxanthohumol derived from hops (Patent Document 1) and the like have been known to have the function of promoting the production of nitric oxide, but it can be considered that in addition to this, nitric oxide-producing components derived from food are also required.

[0004] On the other hand, for polygonatum sibiricum and fennel, their use as a composition for stabilizing blood vessels or strengthening the barrier function of blood vessels has been described (Patent Document 2).

[0005] However, there has been no report on the relationship between polygonatum sibiricum, fennel and the action of promoting nitric oxide production or improving vascular endothelial function.

[0006] Prior Art Documents

[0007] Patent Documents

[0008] Patent Document 1: Japanese Patent No. 7352570

[0009] Patent Document 2: Japanese Unexamined Patent Application Publication No. 2023-97354 Summary of the Invention

[0010] The present invention provides a composition for promoting the production of nitric oxide.

[0011] The inventors of the present invention have found that polygonatum sibiricum and / or fennel are useful for promoting the production of nitric oxide in vascular endothelial cells. The present invention is made based on this insight.

[0012] According to the present invention, the following inventions are provided.

[0013] (1) A composition for promoting nitric oxide production, comprising polygonatum sibiricum and / or fennel.

[0014] (2) The composition according to (1), wherein the promotion of nitric oxide production is carried out in vascular endothelial cells.

[0015] (3) A composition for improving vascular endothelial function, comprising polygonatum sibiricum and / or fennel.

[0016] (4) The composition according to any one of (1) to (3), wherein the polygonatum sibiricum and / or fennel comprises at least one part selected from the group consisting of fruits, rhizomes, flowers, seeds, buds, leaves, aerial stems, and roots.

[0017] (5) The composition according to any one of (1) to (4), which is for oral administration.

[0018] (6) The composition according to any one of (1) to (5), wherein the aforementioned composition is a food or a food additive.

[0019] (7) The composition according to any one of (1) to (6), wherein the aforementioned composition is a health functional food.

[0020] (8) The composition according to any one of (1) to (7), wherein the aforementioned composition is at least one selected from the group consisting of Foods for Specified Health Uses (FOSHU), Foods with nutrient function claims (FNFC), and Foods with function claims (FFC).

[0021] (9) Use of polygonatum sibiricum and / or fennel in the manufacture of a composition for promoting nitric oxide production.

[0022] (10) Use of polygonatum sibiricum and / or fennel in the manufacture of a composition for improving vascular endothelial function.

[0023] (11) A method for promoting nitric oxide production, comprising: administering an effective amount of polygonatum sibiricum and / or fennel to an object that requires it or causing an object that requires it to ingest it.

[0024] (12) A method for improving vascular endothelial function, comprising: administering an effective amount of polygonatum sibiricum and / or fennel to an object that requires it or causing an object that requires it to ingest it.

[0025] (13) Polygonatum sibiricum and / or fennel for promoting nitric oxide production.

[0026] (14)Polygonatum sibiricum and / or fennel for improving vascular endothelial function

[0027] According to the present invention, a composition for promoting nitric oxide production can be provided. In addition, according to the present invention, a composition for improving vascular endothelial function can be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a graph showing a comparison of the amount of nitric oxide produced in human coronary artery endothelial cells when Polygonatum sibiricum or fennel is added. DETAILED DESCRIPTION OF THE INVENTION

[0029] According to one aspect of the present invention, a composition for promoting nitric oxide production is provided. The composition for promoting nitric oxide production contains Polygonatum sibiricum and / or fennel. It should be noted that in this specification, "Polygonatum sibiricum and / or fennel" means Polygonatum sibiricum, fennel, or a combination of Polygonatum sibiricum and fennel.

[0030] According to another aspect of the present invention, a composition for improving vascular endothelial function is provided. The composition for improving vascular endothelial function contains Polygonatum sibiricum and / or fennel.

[0031] "Polygonatum sibiricum" of the present invention is a plant of the genus Polygonatum in the family Liliaceae, also known as Polygonatum falcatum, and is native to Japan and East Asia.

[0032] As the part of Polygonatum sibiricum used in the present invention, for example, fruits, rhizomes, flowers, seeds, buds, leaves, aerial stems, and roots can be used, and the rhizome is preferably used.

[0033] For the Polygonatum sibiricum used in the present invention, both raw Polygonatum sibiricum and dried Polygonatum sibiricum can be used. According to an embodiment of the present invention, "Polygonatum sibiricum" is a Polygonatum sibiricum rhizome or a dried product thereof, preferably a rhizome powder obtained by pulverizing the dried rhizome.

[0034] When Polygonatum sibiricum (including rhizomes, dried powders of rhizomes, extracts, etc.) is commercially available, as long as a substance having the target effect of the present invention can be obtained, it can also be used.

[0035] "Fennel" of the present invention is a plant of the genus Foeniculum in the family Apiaceae, also known as Foeniculum vulgare, and is native to the Mediterranean coastal areas of Europe.

[0036] As the part of fennel used in the present invention, for example, fruits (pulp, pericarp, etc.), rhizomes, flowers, seeds, buds, leaves, aerial stems, and roots are used, and fruits are preferably used.

[0037] For the fennel used in the present invention, either raw fennel or dried fennel can be used. According to an embodiment of the present invention, "fennel" is fennel fruit or its dried product, preferably fruit powder obtained by pulverizing the dried product of the aforementioned fruit.

[0038] When fennel (including fruits, dried powders of fruits, extracts, etc.) is commercially available, as long as a substance having the target effect of the present invention can be obtained, it can also be used.

[0039] Nitric oxide (NO) is synthesized in vivo from arginine using nitric oxide synthase (NOS). Here, nitric oxide dilates blood vessels due to the relaxation of vascular smooth muscle through information transmission systems such as the activation of guanylate cyclase, the increase in cyclic guanosine monophosphate (cGMP), the release of calcium ions to the outside of the cell, and the relaxation of vascular smooth muscle, thereby exerting an antihypertensive effect and being able to prevent and improve hypertension. In addition, nitric oxide exerts an effect of improving vascular endothelial function through the inhibitory effect on platelet aggregation achieved by promoting the production of prostaglandin I2 and the effect of inhibiting the expression of cell adhesion factors that cause thrombosis, and can prevent and improve arteriosclerosis.

[0040] Vascular endothelial cells undertake functions such as maintaining / regulating fluidity and regulating cell movement to maintain vascular constancy. Among these functions, the function of maintaining / regulating fluidity is achieved not only through the regulatory function of vascular endothelial cells on vascular relaxation / constriction, but also through the function of regulating blood coagulation by releasing substances that promote / inhibit blood coagulation. In addition, the regulation of cell movement is achieved through functions such as regulating the expression of vascular adhesion factors.

[0041] The "improvement of vascular endothelial function" of the present invention can be non-invasively measured and diagnosed by flow-mediated dilation (FMD) detection, plethysmography, etc., and preferably promotes the production of nitric oxide in vascular endothelial cells.

[0042] According to a preferred embodiment of the present invention, the promotion of nitric oxide production in the present invention is carried out in vascular endothelial cells. As the vascular endothelial cells of the present invention, there is no limitation. For example, cells obtained from an organism, their passaged cells, or cell line-derived cells can be used, and these cells can also be differentiated and used as needed. The vascular endothelial cells of the present invention can be cells derived from any animal, preferably human cells. Specifically, examples of the vascular endothelial cells of the present invention include: human renal glomerular endothelial cells (HRGEC), human umbilical vein endothelial cells (HUVEC), human aortic vascular endothelial cells (HAEC), human pulmonary artery vascular endothelial cells (HPAEC), human coronary artery vascular endothelial cells (HCAEC), human iliac artery endothelial cells (HIAEC), human microvascular endothelial cells (HMVEC), etc.

[0043] The composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention can be applied both in vivo and in vitro. When the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention are applied to a subject, there is no limitation on the application subject. For example, it includes: primates including humans and chimpanzees; pet animals such as dogs and cats; livestock animals such as cows, horses, sheep, and goats; mammals such as rodents such as mice and rats, etc.

[0044] The compounding amounts of polygonatum and / or fennel in the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention can be appropriately determined according to the use and the intake amount described later. For this compounding amount, for example, as the dry mass of the rhizome of polygonatum or the dry mass of the fruit of fennel relative to the recommended daily intake amount of the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention, examples include 1 mg to 5000 mg, preferably 1 mg to 1000 mg, and more preferably 1 mg to 500 mg. When the composition of the present invention contains both polygonatum and fennel, the aforementioned compounding amount represents their total amount.

[0045] The composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention can be an oral composition or a topical composition. It is considered that polygonatum and / or fennel of the present invention are preferably orally ingested and absorbed by the digestive tract, so that their active ingredients exert effects in the body. Therefore, according to a preferred embodiment of the present invention, the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention are oral compositions.

[0046] In the case where the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention are made into an oral composition, known ingredients (additives) incorporated in the oral composition can be further incorporated. Such ingredients are not limited, and examples include: sweeteners, colorants, preservatives, thickeners, stabilizers, gelling agents, pastes, antioxidants, color developers, bleaching agents, mold inhibitors (antibacterial agents), yeast nutrients (yeast food), gum bases, alkaline noodles, bitter agents, enzymes, brighteners, edible flavors, acidulants, chewing gum softeners, softeners, seasonings, coagulants for tofu, emulsifiers, pH adjusters, bulking agents, nutritional fortifiers and others (manufacturing agents, etc.), food materials, and the like.

[0047] The oral composition of the present invention can be any one of liquid, semi-solid, and solid.

[0048] In the case where the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention are made into a topical composition, known ingredients (additives) incorporated in the topical composition can be further incorporated. Such ingredients are not limited, and examples include: emulsifiers, hydrating agents, solvents, emollients, stabilizers, thickeners, preservatives, lubricants, chelating agents, fillers, excipients, powders, fragrances, edible flavors, absorbents, dyes, opacifiers, antioxidants, vitamins, amino acids, and the like.

[0049] The topical composition of the present invention can be any one of liquid, semi-solid, and solid.

[0050] The composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention can also be any one of pharmaceutical compositions, cosmetic compositions, food compositions (such as food and beverages, food additives), and are preferably food compositions.

[0051] The composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention can also be health functional foods such as foods for specified health uses, foods with nutrition function claims, foods with function claims, etc. Additionally, they can be manufactured in the form of therapeutic diets (i.e., diets that serve a therapeutic purpose, or diets cooked based on a recipe prepared by a dietitian or the like in accordance with a dietary prescription formulated by a doctor), dietary therapy diets, nursing diets, and the like.

[0052] The intake amount of polygonatum and / or fennel per meal or per day in the composition for promoting nitric oxide production and the composition for improving vascular endothelial function of the present invention can be appropriately set according to the object for promoting nitric oxide production, the age, weight, gender of the object for improving vascular endothelial function, or the index values related to nitric oxide and vascular endothelial function before intake, etc., and based on non-clinical or clinical trial results, etc. The intake period of the composition of the present invention can also be appropriately set, and it is preferably repeated long-term and taken daily. In one embodiment of the present invention, as the intake amount of the active ingredient contained in the composition of the present invention (also referred to as the effective amount of polygonatum and / or fennel), for example, 0.001 to 1000 mg / kg body weight / day can be cited, preferably 0.005 to 100 mg / kg body weight / day, and more preferably 0.01 to 10 mg / kg body weight / day. In addition, this intake amount is preferably taken in such a way that it is 5 g / day or less, more preferably 2 g / day or less, and even more preferably 1 g / day or less. The number of administrations can be cited, for example, 1 to 5 times a day, preferably 1 to 3 times a day.

[0053] The use of "promoting nitric oxide production" or "improving vascular endothelial function" of the composition of the present invention can be labeled directly or indirectly. Examples of direct labeling are the descriptions on tangible objects such as the product itself, packaging, container, label, hangtag, etc. Examples of indirect labeling are advertising / promotion activities achieved through places or means such as websites, stores, exhibitions, billboards, bulletin boards, newspapers, magazines, television, radio, mailings, emails, etc. When the composition of the present invention is manufactured in the form of a food with a functional claim, etc., the functionality can be labeled based on the action effects of the ingredients contained as functional ingredients, such as "improving vascular endothelial function", "preventing the reduction of vascular endothelial function", "maintaining the health of blood vessels", "maintaining vascular softness", or equivalent labeling or functional labeling, etc.

[0054] In addition, according to another aspect of the present invention, there is provided the use of polygonatum and / or fennel in the manufacture of a composition for promoting nitric oxide production. In addition, according to yet another aspect of the present invention, there is provided the use of polygonatum and / or fennel in the manufacture of a composition for improving vascular endothelial function.

[0055] According to another aspect of the present invention, there is provided a method for promoting nitric oxide production, which comprises administering an effective amount of polygonatum sibiricum and / or fennel to a subject in need thereof or causing the subject in need thereof to ingest the same. According to still another aspect of the present invention, there is provided a method for improving vascular endothelial function, which comprises administering an effective amount of polygonatum sibiricum and / or fennel to a subject in need thereof or causing the subject in need thereof to ingest the same. The above methods can be either therapeutic or non-therapeutic methods. When administering or causing a subject to ingest polygonatum sibiricum and / or fennel, the production of nitric oxide in vascular endothelial cells is promoted, and the amount of nitric oxide produced increases. Through this effect, vascular endothelial function can be improved.

[0056] In addition, according to another aspect of the present invention, there is provided polygonatum sibiricum and / or fennel for promoting nitric oxide production. In addition, according to still another aspect of the present invention, there is provided polygonatum sibiricum and / or fennel for improving vascular endothelial function.

[0057] In addition, according to another aspect of the present invention, there is provided the use of a composition comprising polygonatum sibiricum and / or fennel for promoting nitric oxide production. According to still another aspect of the present invention, there is provided the use of a composition comprising polygonatum sibiricum and / or fennel for improving vascular endothelial function.

[0058] The above uses in the manufacture, the method for promoting nitric oxide production, the method for improving vascular endothelial function, polygonatum sibiricum and / or fennel, and the uses of the composition comprising polygonatum sibiricum and / or fennel can all be implemented according to the descriptions of the compositions and methods of the present invention.

[0059] Examples

[0060] The present invention will be specifically described based on the following examples, but the present invention is not limited to these examples. Unless otherwise specified, the content represents mass%.

[0061] Preparation of buffer solution, etc.

[0062] HEPES buffer was prepared as follows. 0.22 g of KCl, 7.7 g of NaCl, 0.14 g of Na2HPO4, 1.8 g of D-glucose, and 7.15 g of HEPES (Dojindo Laboratories) were added to ultrapure water and mixed, adjusted to pH 7.4, and then made up to 1 L with ultrapure water to obtain HEPES buffer.

[0063] The 2× recording medium was prepared as follows. 0.80 g of KCl, 13.44 g of NaCl, 0.40 g of CaCl2, 0.152 g of MgCl2, 4.98 g of D-glucose, and 9.54 g of HEPES were added to ultrapure water and mixed, adjusted to pH 7.4 (25 °C) using 4 mol / l KOH, and then ultrapure water was added to make the total volume 1 L to obtain the 2× recording medium. Then, it was filter-sterilized with a 0.2 μm sterilizing filter and stored refrigerated (4 °C). When in use, it was diluted with ultrapure water to 1× recording medium before use.

[0064] The loading buffer (final concentration 2.5 μM DAF-FM DA) was prepared as follows. 5 mL of 2× recording medium, 50 μL of 1.25 mM probenecid, 500 μL of 50 μM DAF-FM DA (Diaminofluorescein-FM diacetate) (Goryo Pharmaceutical Co., Ltd.), and 4.95 mL of ultrapure water were mixed to obtain the loading buffer.

[0065] Preparation of test substance sample applied to endothelial cells

[0066] As the test substance 1 (positive control), adenosine triphosphate disodium (ATP) (FUJIFILM Wako Pure Chemical Corporation) was used. ATP was mixed with HEPES buffer to obtain the test substance 1 sample (n = 4). Additionally, as the test substance 2, the powder of fennel fruit (sterilized product) (Nippon Powdered Drug Co., Ltd.) was used. The powder of fennel fruit was mixed with HEPES buffer to obtain the test substance 2 sample (n = 4). Additionally, as the test substance 3, the powder of the rhizome of Polygonatum sibiricum (sterilized product) (Nippon Powdered Drug Co., Ltd.) was used. The powder of the rhizome of Polygonatum sibiricum was mixed with HEPES buffer to obtain the test substance 3 sample (n = 4).

[0067] Culture of endothelial cells

[0068] The 8th generation of human coronary artery endothelial cells (HCAEC, KURABO) were seeded into 96-well plates and cultured to confluence using HuMedia-EG2 (KURABO). One day before the experiment, the medium was replaced with medium without serum removed from HuMedia-EG2 and cultured overnight. After culturing overnight, the medium was replaced with 100 μL / well of sample loading buffer and cultured at 37 °C and 5% CO2 for 1 hour. After culturing, the supernatant was removed, and 100 μL / well of recording medium was added to prepare the measurement samples. Samples of test substance 1 to test substance 3 were added to the above measurement samples so that the final concentration of test substance 1 reached 10 μM, and the final concentrations of test substances 2 and 3 reached 50 μg / mL, and cultured at 37 °C. After 50 seconds, the fluorescence intensity was measured. It should be noted that only HEPES buffer was added to the control group.

[0069] Measurement of fluorescence intensity

[0070] Using a microplate reader (Infinite200PRO, TECAN), the fluorescence intensity (ex. 485 nm, em. 535 nm) of the measurement samples added with the test substance samples was measured. The obtained results (mean ± standard deviation) are shown in Figure 1 . Regarding the fluorescence intensity of each test substance relative to that of the control group, a significance test for differences between groups (control group vs test substance 1 (ATP) addition group, control group vs test substance 2 (fennel) addition group, control group vs test substance 3 (Polygonatum sibiricum) addition group) was performed by Tukey test (significance level for difference: p < 0.05). As a result, significant differences from the control group were confirmed in the test substance 2 (fennel) addition group and the test substance 3 (Polygonatum sibiricum) addition group. The above results indicate that Polygonatum sibiricum and fennel have the physiological effect of increasing the production of nitric oxide (NO) in vascular endothelial cells (vascular endothelial function improvement effect).

Claims

1. A composition for promoting the production of nitric oxide, comprising polygonatum and / or fennel.

2. The composition according to claim 1, wherein The promotion of nitric oxide production is carried out in vascular endothelial cells.

3. A composition for improving vascular endothelial function, comprising Polygonatum sibiricum and / or Fennel.

4. The composition according to claim 1 or 3, wherein Polygonatum sibiricum and / or fennel include at least one part selected from the group consisting of fruits, underground stems, flowers, seeds, buds, leaves, aerial stems and roots. The composition according to claim 1 or 3, which is for oral administration.

6. The composition according to claim 1 or 3, wherein The composition is a food or beverage or a food additive.

7. The composition according to claim 1 or 3, wherein The composition is a health functional food.

8. The composition according to claim 1 or 3, wherein The composition is at least one selected from the group consisting of foods for specific health purposes, foods with nutritional function claims, and foods with functional claims.

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