Fungal secondary metabolites are considered to be good sources for the screening of lead compounds of clinical drugs. Among fungal species, ~6400 compounds are obtained from various filamentous fungi.1 Among them ~950, 900 and 350 compounds are isolated from Aspergillus, Penicillium and Fusarium species, respectively. Moreover, several other members of filamentous and endophytic genus (Trichoderma, Phoma, Alternaria, Acremonium and Stachybotrys) also produce several hundreds of bioactive compounds. However, the rate of discovery of novel compounds from these fungi has decreased significantly. To increase the chances of discovering novel compounds, we chose to examine rare endophytic fungi. During our screening program, we isolated a novel cytotoxic compound, MBJ-0005 (1), consisting of a highly oxygenated ergostane-based skeleton, from the culture of Anthostomella eucalyptorum f25427. Anthostomella sp. are endophytic fungi, and notably only a few succinic acid derivatives from Anthostomella sp. have been reported.2 Herein, we report the isolation, structure determination and brief biological activity of 1.