New antifungal antibiotics pradimicins D and E were isolated from the culture filtrates of Actinomadura hibisca P157-2 (ATCC 53557) and its mutant A2660 (ATCC 53762). The structure of pradimicin D is N-[[(5S, 6S)-5-O-[4, 6-dideoxy-4-(methylamino)-3-O-(β-D-xylopyranosyl)-β-D-galactopyranosyl]-5, 6, 8, 13-tetrahydro-1, 6, 9, 14-tetrahydroxy-11-methoxy-3-methyl-8, 13-dioxobenzo[a]naphthacen-2-yl]carbonyl]glycine, based on spectral analyses compared to pradimicin A. Pradimicin E is the des-N-methyl analog of pradimicin D. Pradimicins D and E were equal in activity to pradimicin A in vitro against a variety of fungi and in vivo against Candida albicans A9540 in mice. © 1990, JAPAN ANTIBIOTICS RESEARCH ASSOCIATION. All rights reserved.