Highly cytotoxic polypeptides, polytheonamides A-C, have been isolated from the marine sponge Theonella swinhoei. Interpretation of the 2D NMR data of the acid hydrolyzate of polytheonamide B as well as amino acid analysis led to identification of Ala, Asp, Thr, aThr, Ser, Glu, Val, Gly, Ile, t-Leu,beta-methylGlu,beta-methyille,beta-hydroxyVal, and beta-hydroxyAsp. 2D NMR data of polytheonamide B suggested the presence of gamma-hydroxy-t-leu, which was a new amino acid.