Composition containing biochanin a as active ingredient for adenosine a2a receptor activating effect

Abstract

To provide a composition which contains biochanin A as an active ingredient for an adenosine A2A receptor activating effect.SOLUTION: A composition contains biochanin A as an active ingredient for an adenosine A2A receptor activating effect. There are also provided foods with functional claims, supplements, and foods for specified health uses for the purpose of improving the quality of sleep which are obtained using the composition.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition containing biochanin A as an active ingredient that activates adenosine A2A receptors. [Background technology]

[0002] Adenosine A2A receptors are G protein-coupled receptors distributed throughout the body and are known to be deeply involved in sleep regulation in the sleep center of the brain. Activating this adenosine A2A receptor can induce deep non-REM sleep and improve sleep quality. For example, it is known that CGS21680, an adenosine A2A receptor agonist, increases non-REM sleep when administered to animals (see, for example, Non-Patent Documents 1 and 2). As a natural ingredient that activates the adenosine A2A receptor, sake yeast has been reported to increase non-REM sleep in humans and improve sleep quality through activation of the adenosine A2A receptor (see, for example, Patent Document 1, Non-Patent Documents 3 and 4).

[0003] Phytoestrogens including biochanin A are known to enhance immune function (see, for example, Patent Document 2). It is also known that isoflavones containing this compound suppress or reduce an increase in diastolic blood pressure (see, for example, Patent Document 3). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2013-82641 A [Patent Document 2] Special Publication No. 2003-504341 [Patent Document 3] JP 2006-96717 A [Non-patent literature]

[0005] [Non-Patent Document 1] Satoh S, et al. Proceedings of the National Academy of Sciences of the United States of America. 1996 Jun 11;93(12):5980-4. DOI:10.1073 / pnas.93.12.5980. [Non-Patent Document 2] Korkutata M, et al. Neuropharmacology. 2019 Jan;144:122-132. DOI:10.1016 / j.neuropharm.2018.10.022. [Non-Patent Document 3] Nakamura Y, et al. Journal of sleep research. 2016 Dec;25(6):746-753. DOI:10.1111 / jsr.12434. [Non-Patent Document 4] Monoi N, et al. Journal of sleep research. 2016 Feb;25(1):116-23. DOI:10.1111 / jsr.12336. Summary of the Invention [Problem to be solved by the invention]

[0006] The object of the present invention is to provide a substance that activates the adenosine A2A receptor, and a functional food, supplement, or food for specified health uses that uses the substance and is intended to improve sleep quality. [Means for solving the problem]

[0007] As a result of intensive research conducted by the inventors in order to achieve the above-mentioned object, they discovered that biochanin A activates adenosine A2A receptors, thereby improving the quality of sleep, and thus completed the present invention. Therefore, the present invention is a composition containing biochanin A as an active ingredient that activates adenosine A2A receptors. It is also used in functional foods, supplements, and foods for specified health uses that aim to improve sleep quality. Effect of the Invention

[0008] The composition of the present invention containing biochanin A as an active ingredient for activating adenosine A2A receptors is considered to be safe with no reports of adverse events when biochanin A is taken in the amount specified in the product of the present invention. The present invention can increase non-REM sleep and improve sleep quality. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] The present invention will be described in detail below. Biochanin A of the present invention is represented by chemical formula 1 and is an isoflavonoid contained in legumes such as red clover, chickpeas, alfalfa, peanuts, and soybeans, as well as in cabbage, and is known to have effects such as improving lipid metabolism and carbohydrate metabolism via activation of PPARα and PPARγ, as well as estrogen-like effects. When biochanin A is ingested orally, it is absorbed into the body, transferred into the blood, and circulated throughout the body.

[0010] [ka]

[0011] <How to obtain Biochanin A> As mentioned above, biochanin A is contained in the seeds, leaves, stems, and flowers of red clover and other plants, and can be obtained by extraction using methods such as solvent extraction using ethanol, methanol, ethyl acetate, acetone, etc., followed by hydrolysis and purification by column chromatography, etc. Furthermore, biochanin A is commercially available, and commercially available biochanin A can also be used in the present invention.

[0012] <How to use Biochanin A> The method of using Biochanin A is the same as that of conventional Biochanin A and is not particularly limited. For example, it can be incorporated into functional foods, supplements, and foods for specified health uses. Examples of foods that may be designated as functional foods or foods for specified health uses include noodles, rice, bread, Western confectioneries, Japanese confectioneries, chewing gum, juice, and alcoholic beverages. The oral dosage of the composition of the present invention containing biochanin A as an active ingredient having an adenosine A2A receptor activating effect is usually 1 mg to 150 mg / day, preferably 10 mg to 30 mg / day, in terms of the daily intake of biochanin A by a human. EXAMPLES

[0013] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. <Establishment of an adenosine A2A receptor activity evaluation system> The following reporter assay was constructed to evaluate adenosine A2A receptor activity. When adenosine A2A receptors are activated, they interact with intracellular Gs proteins and activate adenylate cyclase, thereby producing cAMP. Activation of adenosine A2A receptors can be evaluated by detecting cAMP signals. The cAMP signal can be detected by measuring the amount of luminescence from firefly luciferase and Renilla luciferase as follows. (1) A plasmid encoding firefly luciferase (product name: pGL4.29, manufactured by Promega Corporation) downstream of CRE (cAMP-response element) and a Renilla luciferase expression vector (product name: pF9A CMV hRluc-neo Flexi® Vector, manufactured by Promega Corporation; Barnase sequence was removed) were introduced into HEK293 cells. A transfection reagent (product name: FuGENE (registered trademark) HD ​​Transfection Reagent, manufactured by Promega Corporation) was used for gene introduction. (2) 5 × 10 cells obtained in (1) were added to a 96-well plate in DMEM medium. 4 Cells were seeded per well. (3) After 23 hours, ZM241385 dissolved in DMSO was added to the samples at the concentrations shown in Samples 1 to 4 below. (4) One hour after (3) above, the medium from (2) above was removed and replaced with DMEM medium containing 10% FBS, and NECA dissolved in DMSO was added to the concentrations shown in Samples 1 to 4 below. (5) Five hours after the medium change in (4) above, the medium was removed, and the luminescence intensities of firefly luciferase and Renilla luciferase were measured using the Dual-Glo (registered trademark) Luciferase Assay System (Promega Corporation). (6) The luminescence intensity of Renilla luciferase was used as a standard, and the luminescence intensity of firefly was compared to this to calculate the ratio. (7) The ratio obtained in (6) above was converted so that the ratio when nothing was added (sample 1) was 1, and the relative amount of light emission was calculated.

[0014] *NECA (5'-N-Ethylcarboxamidoadenosine) is a general adenosine receptor agonist that exhibits high affinity for the adenosine A2A receptor. *ZM241385 is a selective antagonist of the adenosine A2A receptor.

[0015] Sample 1 contained neither NECA nor ZM241385. In sample 2, NECA was added to the medium to a concentration of 100 nM, and ZM241385 was not added. In sample 3, NECA was not added, and ZM241385 was added to the medium to a concentration of 10 μM. In sample 4, NECA was added to the medium to a concentration of 100 nM, and ZM241385 was added to the medium to a concentration of 100 nM.

[0016] As a result, the relative light emission amount of sample 1 was 1.00, the relative light emission amount of sample 2 was 12.17, the relative light emission amount of sample 3 was 1.03, and the relative light emission amount of sample 4 was 1.14. NECA, an agonist of the adenosine A2A receptor, activated the cAMP signal, and ZM241385, an antagonist selective for the adenosine A2A receptor, inhibited the activation of the cAMP signal, demonstrating that the activation of the adenosine A2A receptor can be evaluated using this evaluation system.

[0017] <Adenosine A2A receptor activity evaluation of Biochanin A> The reporter assay was performed in the same manner as above, except that the concentration in the NECA medium was changed from 100 nM to 10 μM in the biochanin A medium. Sample 5 contained neither biochanin A nor ZM241385. In sample 6, biochanin A was added to the medium to a concentration of 10 μM, and ZM241385 was not added. In sample 7, biochanin A was added to the medium to a concentration of 10 μM, and ZM241385 was added to the medium to a concentration of 10 μM.

[0018] As a result, the relative luminescence intensity of sample 5 was 1.00, that of sample 6 was 1.60, and that of sample 7 was 0.99, confirming that biochanin A activates the adenosine A2A receptor.

[0019] [Example 1 Beverage] A beverage was produced by mixing 30 g of fructose glucose liquid sugar, 0.5 g of emulsifier, 1 g of soybean-derived extract containing 1% by mass of biochanin A, an appropriate amount of flavoring, and 60 g of purified water in a mixer in a conventional manner.

Claims

1. A composition containing biochanin A as an active ingredient that activates adenosine A2A receptors.

2. A functional food, supplement, or food for specified health uses intended to improve sleep quality, using a composition containing biochanin A described in claim 1 as an active ingredient with an adenosine A2A receptor activation effect.

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