Three new sulfur-containing alkaloids, polycarpaurines A (1), B (2), and C (3) were isolated from the tropical ascidian Polycarpa aurata collected in Indonesia, together with six known compounds (4–9). The structures of new compounds were assigned on the basis of their spectral data. Compounds 1, 3, 4, and 8 inhibited colony formation of Chinese hamster V79 cells with EC50 values of 6.8, 8.6, 3.8, and 10 mM, respectively. Compounds 2 and 7 showed modest activity against V79 cells (EC50>10 mM).