Paphiopedilum callosum (Rchb.f.) Stein belongs to the Orchidaceae family. The phytochemistry and bioactivities of this plant have not been reported. In this study, a bioassay-guided isolation with a yeast cell-based assay was performed to isolate topoisomerase I poison compounds from the roots of P. callosum. One new and five known compounds were isolated. According to their spectroscopic data (ESI-MS, NMR, IR and UV), the new compound (1) is 3′-hydroxy-2,6,5′-trimethoxystilbene. The known compounds (2-6) were identified by comparing their spectroscopic data with those in the literature. According to the yeast cell-based assay, all the compounds were topoisomerase Ι poisons based on their ability to inhibit the growth of yeast cells. As further confirmation, the cytotoxic activities of the isolated compounds were evaluated using human cancer cell lines (MCF-7 and NCI-H187). The compounds exhibited varying degrees of cytotoxicity on the representative cell lines. These isolated compounds could serve as new lead compounds in the development of cancer chemotherapeutics.
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