A new prenylated indole alkaloid, neofipiperzine D, was isolated from the cultures of Neosartorya fischeri, together with three known metabolites fumitremorgin C, ergosterol, and ergosterol peroxide. The structure of the new compound was established by detailed analysis of its spectroscopic data, especially 1D, 2D NMR and HR-ESI-MS data. In an in vitro assay, neofipiperzine D did not exhibit any significant cytotoxicity against MCF-7, H1299, HUVEC, and MDA-MB-231 cell lines at a concentration of 20 μM. © 2014, Science Reviews 2000 Ltd. All rights reserved.