Rhizovagine A (1), a novel dibenzo-alpha-pyrone alkaloid with an unprecedented 5/5/6/6/6 fused pentacyclic skeleton, was isolated from the endophytic fungusRhizopycnis vagumNitaf22. The structure was elucidated by comprehensive spectroscopic analysis, in combination with quantum chemical(13)C NMR and electronic circular dichroism (ECD) calculations for configurational assignment. A plausible biosynthetic pathway for1was proposed. Compound1displayed acetylcholinesterase inhibitory activity.