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

Abstract

To provide a composition containing rutaecarpine as an active ingredient for the activation of adenosine A2A receptor.SOLUTION: Provided is a composition containing rutaecarpine 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 rutaecarpine 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] Rutaecarpine is known to inhibit cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2) (see, for example, Patent Document 2). It is also known to have a cardiotonic effect, such as cardiovascular protection, via TRPV1 (see, for example, Non-Patent Document 5). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2013-82641 A [Patent Document 2] Special Publication No. 2022-505857 [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] Sujie Jia, et al. Molecules. 2010 Mar 15;15(3):1873-81. DOI:10.3390 / molecules15031873. 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 rutaecarpine activates adenosine A2A receptors, thereby improving sleep quality, and thus completed the present invention. Therefore, the present invention is a composition containing rutaecarpine 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 rutaecarpine as an active ingredient with adenosine A2A receptor activating effect is considered to be safe with no reports of adverse events when rutaecarpine is taken at the intake 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 rutaecarpine of the present invention is represented by chemical formula 1 and is contained in Euonymus chinensis, a citrus plant native to China, as well as some Zanthoxylum genus plants. Euonymus chinensis is a raw material for traditional Chinese medicine used for headaches, stomachaches, etc., and rutaecarpine is known to have cardiotonic, vasodilatory, and anti-inflammatory effects. When ingested orally, rutaecarpine is absorbed into the body, enters the bloodstream, and circulates throughout the body.

[0010] [ka]

[0011] <How to get Rutaecarpine> As mentioned above, rutaecarpine is contained in the fruit of the Rutaceae plant Euonymus chinensis and some species of the Zanthoxylum genus, 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. Rutaecarpine is commercially available, and commercially available rutaecarpine can also be used in the present invention.

[0012] <How to use Rutaecarpine> The method of using rutaecarpine is the same as that of conventional rutaecarpine 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 rutaecarpine as an active ingredient having adenosine A2A receptor activation activity is typically 1 mg to 1000 mg / day, preferably 1 mg to 500 mg / day, and more preferably 1 mg to 100 mg, in terms of the daily human rutaecarpine intake. 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] <Adenosine A2A receptor activity evaluation of rutaecarpine> 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 rutaecarpine medium. Sample 5 contained neither rutaecarpine nor ZM241385. In sample 6, rutaecarpine was added to the medium to a concentration of 10 μM, and ZM241385 was not added. In sample 7, rutaecarpine 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, the relative light emission intensity of sample 6 was 2.05, and the relative light emission intensity of sample 7 was 1.19, confirming that rutaecarpine 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 an extract derived from Euonymus chinensis containing 1% by mass of rutaecarpine, an appropriate amount of flavoring, and 60 g of purified water in a mixer in a conventional manner.

Claims

1. A composition containing rutaecarpine 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 the rutaecarpine described in claim 1 as an active ingredient with an adenosine A2A receptor activation effect.

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