The structure of a new cytotoxic macrolide, superstolide B [1], isolated from the deep water sponge Neosiphonia superstes, collected off New Caledonia, was elucidated mainly on the basis of nmr data. Compound 1 is closely related to superstolide A [2], a major cytotoxic component isolated from that organism, but lacks the 25-hydroxyl group found in 2 and has a C-24 (C-25)-double bond.