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A new bicyclic nonene, tsaokoic acid (<b>1</b>), was isolated from the fruits of <i>Amomum tsao-ko</i>, together with three known compounds (<b>2</b>–<b>4</b>). The structure of <b>1</b> was elucidated by analyzing spectroscopic data including 1D and 2D NMR spectra and compounds <b>2</b>–<b>4</b> were identified as tsaokoin, vanillin, and tsaokoarylone, respectively, by comparing their NMR spectra with previously reported data. Compounds <b>1</b>–<b>4</b> showed possible inhibitory activity against acetylcholinesterase (AChE) in silico molecular docking simulations. They were submitted to in vitro assay system and exhibited moderate inhibitory activity with IC₅₀ values of 32.78, 41.70, 39.25, and 31.13 μM, respectively.