Among fungi, the genera Penicillin, Aspergillus, Trichoderma and Fusarium have appeared in a considerable number of reports as producers of not only antimicrobial agents but also a variety of bioactive compounds. We are attempting to develop resources for drug discovery by characterizing previously unknown fungi or even known fungi, which have been rarely reported to produce secondary metabolites. Fungal strain FKA-25 was isolated from a soil sample collected at Yakushima Island, Kagoshima Prefecture. FKA-25 was considered to belong to the genus Pseudobotrytis based on taxonomic characteristics. There have been some reports on this genus, but no reports on the secondary products. Sespendole was purified from EtOAc extracts of the 4 day-old culture broth of the strain by ODS chromatography and HPLC. The molecular formulae was determined to be C_<33>H_<45>O_4N by HR-EI-MS. The structure was elucidated by various NMR experiments. Sespendole possesses the indole-sesquiterpene core with two additional isoprenyl side chains. To our knowledge, sespendole is the first microbial metabolite having the indole-sesquiterpene skeleton. The relative stereochemistries of indole-sesquiterpene moiety are presumed to be 3S^*, 4R^*, 9S^* and 12S^* from the NOE experiments. The biosynthetic origin of sespendole was studied by feeding the kinds of sodium [^<13>C]acetates to the strain, indicating that the indole-sesquiterpene core was derived from a sesquiterpene and an indole ring. Sespendole inhibited the synthesis of cholesteryl ester and triacylglycerol by mouse macrophages with IC_<50> values of 4.0 and 3.2μM, respectively.