Sulfated steroids have attracted a great attention by their unusual chemical structures,1 activity against HIV,2-7 and as inhibitors of enzymes8, 9 and have been described from a wide variety of marine organisms, particularly sponges and echinoderms. As part of our continuing search for biologically active natural products from marine organisms, it was found that crude extract of an unidentified marine ascidian exhibited potent cytotoxic activity. Bioassay-guided fractionation of the extract yielded new sulfated streoids and their structures were determined on the basis of spectroscopic data. These compounds exhibited cytotoxicity against human leukemia cell line (K-562).