The three new 10-phenyl-[12]-cytochalasins Z7, Z8, and Z9 (1-3), together with two known analogues, cytochalasins E (4) and K (5), were isolated from the marine-derived fungus Spicaria elegans. This is only the second report to date that cytochalasins contain a 12-membered macrocyclic ring. The structures of the three new cytochalasins were elucidated by spectroscopic methods, and their absolute configurations were determined for the first time by the modified Mosher's method. All five cytochalasins were evaluated for their cytotoxic effects on P388 and A-549 cell lines by the MTT method.