Composition containing apigenin as active ingredient for activation of adenosine a2a receptor

Abstract

To provide a composition containing apigenin as an active ingredient for the activation of adenosine A2A receptor.SOLUTION: Provided is a composition containing apigenin as an active ingredient for the activation of adenosine A2A receptor. Furthermore, provided is a functional food with health claims, a supplement, or a food for specified health uses, intended for improving sleep quality by using the composition.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition containing apigenin 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] Apigenin is known to have an inhibitory effect on microglial activation (see, for example, Patent Document 2). It is also known to have a sedative effect via the benzodiazepine binding site of the GABA A receptor, and an antidepressant effect via serotonin, dopamine, and α-adrenergic transmission (see, for example, Non-Patent Documents 5 and 6). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2013-82641 A [Patent Document 2] Special Publication No. 2020-183346 [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. [Non-Patent Document 5] Viola H, et al. Planta Medica. 1995 Jun;61(3):213-6. DOI: 10.1055 / s-2006-958058. [Non-Patent Document 6] Al-Yamani MJ,et al. Saudi Journal of Biological Science. 2022 Jan;29(1):11-17. DOI: 10.1016 / j.sjbs.2021.11.008. [Summary of the Invention] [Problems 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 apigenin activates adenosine A2A receptors, thereby improving the quality of sleep, and thus completed the present invention. Therefore, the present invention is a composition containing apigenin 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 apigenin as an active ingredient with adenosine A2A receptor activating effect is considered to be safe with no reports of adverse events, when apigenin 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. The apigenin of the present invention is represented by Chemical Formula 1, and is contained in chamomile, guava, Lamiaceae plants such as thyme, oregano, peppermint, etc., and Apiaceae plants such as celery, parsley, angelica tree, cumin, etc. Apigenin is known to have sedative, anti-anxiety, anti-depressant, etc. When apigenin is orally ingested, it is absorbed into the body, transferred into the bloodstream, and circulated throughout the body.

[0010] [ka]

[0011] <How to obtain apigenin> As described above, apigenin is contained in the leaves, stems, roots, flowers, seeds, fruits, etc. of plants such as chamomile, guava, thyme, and celery, and can be obtained by extraction using a method such as solvent extraction using ethanol, methanol, ethyl acetate, acetone, etc., followed by hydrolysis and purification by column chromatography, etc. Apigenin is also commercially available, and commercially available apigenin can also be used in the present invention.

[0012] <How to use Apigenin> The method of using apigenin is the same as that of conventional apigenin 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 apigenin as an active ingredient having adenosine A2A receptor activation activity is usually 1 mg to 5000 mg / day, preferably 1 mg to 500 mg / day, and more preferably 10 mg to 100 mg, in terms of the daily apigenin intake 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 10 μM.

[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] <Analysis of apigenin's adenosine A2A receptor activity> The reporter assay was performed in the same manner as above, except that the NECA concentration in the medium was changed from 100 nM to 10 μM in the apigenin medium. Sample 5 contained neither apigenin nor ZM241385. In sample 6, apigenin was added to the medium to a concentration of 10 μM, and ZM241385 was not added. In sample 7, apigenin 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 light emission intensity of sample 5 was 1.00, that of sample 6 was 2.00, and that of sample 7 was 1.08, confirming that apigenin 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 chamomile-derived extract containing 1% by mass of apigenin, an appropriate amount of flavoring, and 60 g of purified water in a mixer in a conventional manner.

Claims

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

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

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