Evolving the structural architecture of prodiginines for next-generation antimalarial development and target identification - proposal summary despite modern intervention, malaria has endured as a…

Evolving the structural architecture of prodiginines for next-generation antimalarial development and target identification - proposal summary despite modern intervention, malaria has endured as a leading cause of morbidity and mortality worldwide, resulting in around 250 million cases and 600,000 deaths annually. The emergence and escalation of drug resistance in the causative plasmodium parasites, including to the most widely used artemisinin-based combination therapies, underscores the urgent need for new therapeutics with alternative mechanisms of action that can circumvent this resistance. The prodiginines, a class of microbial natural products characterized by a core tripyrrole scaffold, exhibit potent antimalarial activity and low cytotoxicity, positioning them as promising leads in the discovery and development of novel antimalarial drugs. However, the full scope of their therapeutic potential has yet to be realized, largely due to the structural complexity of prodiginine scaffolds. While various approaches-including total synthesis, fermentation, and mutagenesis-have been employed to generate and modify prodiginines, these methods alone have proven insufficient to comprehensively explore the full architectural landscape of this natural product class. Hence, this proposal seeks to address these limitations by developing semi-synthetic modification and DE novo combinatorial biosynthesis strategies for the production and expansion of prodiginine scaffolds. Aim 1 leverages late-stage semi-synthetic pyrrole functionalization to produce derivatives of premarineosin a, a cyclic prodiginine with potent (EC50 < 5 NM) antimalarial activity against drug-sensitive and drug-resistant plasmodium falciparum. Using this same chemistry to attach chemical handles to the premarineosin a scaffold will enable pull-down proteomics to identify its protein targets and elucidate its mode of action. Aim 2 harnesses metabolic engineering of combinatorial biosynthetic pathways to produce non- natural prodiginine scaffolds toward potentially novel modes of antimalarial activity. The design of a modular chassis strain will enable the generation of tailored hybrid prodiginines via a plug-and-play approach. Collectively, these efforts will yield an expanded library of prodiginine analogs, provide new insights into their molecular targets, and design versatile biosynthetic strategies for scalable production of prodiginine-like scaffolds. By systematically expanding access to unique prodiginine analogs, this proposal will accelerate the discovery of novel antimalarial therapeutics that can overcome the ongoing resistance crisis. Moreover, this work will be conducted within a dynamic and collaborative research environment that provides comprehensive mentorship and interdisciplinary training opportunities, developing expertise at the interface of synthetic chemistry, microbial engineering, and translational drug discovery and promoting success in an independent academic career.