Eleven previously undescribed arylnaphthalide lignans (<b>1</b>-<b>11</b>) together with seven known compounds were isolated from the whole plant of <i>Justicia depauperata</i>. The structures of <b>1</b>-<b>11</b> were elucidated by spectroscopic analysis and mass spectrometry. Compounds <b>6</b> (IC<sub>50</sub> = 4.1 μM) and <b>9</b> (IC<sub>50</sub> = 9.5 μM) displayed cytotoxic activity against the KB-3-1 cervical carcinoma cell line. This report provides an insight into the conformational equilibria occurring in the arylnaphthalide lignan constituents of this plant.