John M. Billingsley, Jiaming Ding, Allison T. Hands, Kanji Niwa, Nathan J. Adamson, Lukas A. Wein, Bruno Perlatti, Neil K. Garg, Yi Tang
Interrupted reactions, in which an intermediate is redirected from its conventional mechanistic pathway, offer a unique approach to the assembly of complex natural products. This study details the biosynthesis of a newly discovered family of alkaloids named the subrubines alongside the total synthesis of the penultimate member, pensubrubine, featuring distinct interrupted reaction pathways. These natural products, identified using high-resolution genome mining of an active site mutation, represent the only reported microbial diaza[3.3.3]-propellane pyrrolidinoindolines. We demonstrate through complete pathway reconstitution that the putatively annotated ene-reductase SubF functions as the propellane synthase that directs an enolate intermediate toward an intramolecular Mannich cyclization. Concurrently, a concise 7-step total synthesis of pensubrubine was developed, employing a diastereoselective interrupted Fischer indolization reaction to rapidly construct the diaza[3.3.3]-propellane core and establish the absolute configuration of pensubrubine. The reported bio- and total syntheses of subrubines showcase the value of interrupted pathways in assembling complex scaffolds.