In this paper we report the results of the study of the in vitro effect of eight anticancer DNA topoisomerase inhibitors on the growth of Aspergillus fumigatus, A. niger, Candida glabrata and Cryptococcus neoformans. Only one compound, idarubicin, displayed promising antifungal activity against A. niger, C. glabrata and C. neoformans with MIC(50) values varying between 3.6 and 14.2 μM (1.8-7.1 μg/ml). Three other compounds, aclarubicin, doxorubicin and mitoxantrone, showed some antifungal activity against C. glabrata and C. neoformans with MIC(50) values in the mid micromolar range. The data of this study indicate that selected DNA topoisomerase inhibitors are a promising class of compounds for the development of new antifungal agents.