Isolation and Structure Elucidation of Bioactive Constituents from the Fruits of Amomum tsaoko with AChE Inhibitory Activity

Abstract

Amomum tsao-ko Crevest et Lemarie (생강과: Zingiberaceae)는 중국에서 사용되는 약용식물로 여러가지 약리활성과 독특한 향으로 약재와 향신료로 사용되어져 왔다. A. tsaoko의 열매는 복부팽만, 구토, 설사, 말라리아에 사용되어져 왔다고 보고하고 있다. A. tsaoko는 향신료로의 활용은 많이 보고가 되어있으나 다른 약재에 비해 구성물질 및 활성이 많이 보고되어 있지 않다. A. tsaoko의 열매와 씨를 MeOH로 추출하여 이를 Hexane, Ethyl acetate, Butanol로 분획을 진행하여 나누었다. 이중 Ethyl acetate 분획물을 Column chromatography와 MPLC, HPLC를 통해 단일성분인 tsaokoarylone (7), 8-hydroxy-2,6-dimethyl-2,6-octadienal (18), vanillin (22), tsaokoin (28), tsaokoic acid (31)인 총 5개의 물질을 분리했다. 분리된 단일 성분은 1D NMR과 2D NMR 분석을 통해 구조를 확인했고 이전에 분리되어 보고되었던 tsaokoarylone (7), 8-hydroxy-2,6-dimethyl-2,6-octadienal (18), vanillin (22)의 구조를 다른 문헌과 비교하여 확인했다. 이후 활성을 확인하기위해 in silico docking simulation과 in vitro 실험을 진행했고 이를 통해 acetylcholinesterase (AChE) 억제에 대해 중간 활성을 보임을 확인했다. 결과적으로 A. tsaoko가 acetylcholinesterase (AChE)를 억제하는 활성이 존재함을 밝혔으며 이전에 분리되지 않았던 tsaokoic acid (31)를 분리하여 구조를 규명했다. ;Amomum tsao-ko Crevest et Lemarie (Gingeraceae: Zingiberaceae) is a medicinal plant used in China. It has been used as a medicinal and spice due to its various pharmacological actions and unique flavor. It has been reported that the fruits of A. tsaoko have been used for bloating, vomiting, diarrhea and malaria. Although it has been reported that A. tsaoko is used as a spice, the composition and activity of A. tsaoko have not been reported as much compared to other herbal medicines. The fruits and seeds of A. tsaoko were extracted with MeOH and fractionated with Hexane, Ethyl acetate and Butanol. Then, the ethyl acetate fraction was separated by column chromatography, MPLC and HPLC to separate five substances into single components, tsaokoarylone (7), 8-hydroxy-2,6-dimethyl-2,6-octadienal (18), vanillin (22), tsaokoin (28), tsaokoic acid (31). The structures of the 5 isolated single components were confirmed by 1D NMR and 2D NMR, and the structures of tsaokoarylone (7), 8-hydroxy-2,6-dimethyl-2,6-octadienal (18), vanillin (22), tsaokoin (28), previously reported separately were confirmed by comparison with other literatures. As a result of performing in silico docking simulation and in vitro assay to confirm the subsequent activity, it was confirmed that the moderate activity of acetylcholinesterase (AChE) inhibitory was shown. As a result, it was revealed that A. tsaoko had acetylcholinesterase (AChE) inhibitory activity, and the structure was elucidated by identification tsaokoic acid (31), which had not been previously isolated.