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A new tetrahydrofuran lignan, (7S,8R,7'S,8'S)-3-methoxy-3',4'-methylenedioxy-7,9'-epoxylignane-4,7',9-triol (1), and 21 known compounds 2–22 were isolated from the roots of Asiasarum heterotropoides by chromatographic separation methods. The structures of all compounds 1–22 were elucidated by spectroscopic analysis including 1D- and 2D-NMR. Fourteen of these compounds (1–3, 7, 10, 12–17, 19, 21, and 22) were isolated from this species in this study for the first time. All of the isolates were evaluated for their anticancer activities using in vitro assays. Among the 22 tested compounds, two (compounds 5 and 7) induced the downregulation of NO production, FOXP3 expression, and HIF-1α transcriptional activity.