ABSTRACT
Natural products provide an unparalleled source of chemical scaffolds with diverse biological activities and have profoundly impacted antimicrobial drug discovery. To further explore the full potential of their chemical diversity, we survey natural products for antifungal, target-specific inhibitors by using a chemical-genetic approach adapted to the human fungal pathogen Candida albicans and demonstrate that natural-product fermentation extracts can be mechanistically annotated according to heterozygote strain responses. Applying this approach, we report the discovery and characterization of a natural product, parnafungin, which we demonstrate, by both biochemical and genetic means, to inhibit poly(A) polymerase. Parnafungin displays potent and broad spectrum activity against diverse, clinically relevant fungal pathogens and reduces fungal burden in a murine model of disseminated candidiasis. Thus, mechanism-of-action determination of crude fermentation extracts by chemical-genetic profiling brings a powerful strategy to natural-product-based drug discovery.
Subject(s)
Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Biological Products/pharmacology , Biological Products/therapeutic use , Candida albicans/drug effects , Candida albicans/genetics , Drug Evaluation, Preclinical/methods , Polynucleotide Adenylyltransferase/antagonists & inhibitors , Alleles , Amino Acid Sequence , Animals , Antifungal Agents/chemistry , Antifungal Agents/isolation & purification , Aspergillus fumigatus/drug effects , Aspergillus fumigatus/growth & development , Aspergillus fumigatus/metabolism , Biological Products/chemistry , Biological Products/isolation & purification , Candida albicans/metabolism , Candidiasis/drug therapy , Candidiasis/metabolism , Complex Mixtures/pharmacology , Deoxyadenosines/metabolism , Deoxyadenosines/pharmacology , Drug Resistance, Fungal , Fermentation , Heterozygote , Mice , Microbial Sensitivity Tests , Molecular Sequence Data , Mutation , Polyadenylation/drug effects , Polynucleotide Adenylyltransferase/genetics , Polynucleotide Adenylyltransferase/metabolism , RNA, Messenger/metabolism , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/metabolism , Treatment OutcomeABSTRACT
Mechanism-of-action (MOA) studies of bioactive compounds are fundamental to drug discovery. However, in vitro studies alone may not recapitulate a compound's MOA in whole cells. Here, we apply a chemogenomics approach in Candida albicans to evaluate compounds affecting purine metabolism. They include the IMP dehydrogenase inhibitors mycophenolic acid and mizoribine and the previously reported GMP synthase inhibitors acivicin and 6-diazo-5-oxo-L-norleucine (DON). We report important aspects of their whole-cell activity, including their primary target, off-target activity, and drug metabolism. Further, we describe ECC1385, an inhibitor of GMP synthase, and provide biochemical and genetic evidence supporting its MOA to be distinct from acivicin or DON. Importantly, GMP synthase activity is conditionally essential in C. albicans and Aspergillus fumigatus and is required for virulence of both pathogens, thus constituting an unexpected antifungal target.
