Isolation of Natural Prodrug-Like Metabolite by Simulating Human Prodrug Activation in Filamentous Fungus
Chem Pharm Bull (Tokyo). 2022;70(4):304-308. doi: 10.1248/cpb.c21-01099.
Prodrugs have seen increased clinical applications as therapeutic agents, as they reduce undesirable side effects and improve the therapeutic potential of drugs. While microorganisms produce numerous secondary metabolites with useful medicinal properties, there are only a handful of naturally occurring prodrugs discovered to date. The techniques of isolating secondary metabolites with therapeutic potential from natural product producers have been developed extensively over the years. However, the methods of identifying prodrugs from microbes have not been examined in depth, partly because prodrug-type compounds inherently lack the biological activities that are often used to screen for therapeutically useful secondary metabolites. Therefore, we hypothesized that the difficulty in searching for natural prodrug-type compounds may be addressed by simulating human prodrug activation within natural product-producing microbes. We chose to introduce human CYP (hCYP) into natural product-producing filamentous fungi, because hCYPs are the key enzymes that activate prodrugs in human body, and filamentous fungi are known to be prolific producers of a wide variety of natural products. Here, we successfully identified a cytotoxic, antibiotic and potential anti-diabetic natural product leporin B from Aspergillus flavus that was previously not known to produce this compound. Through bioinformatic and metabolite analyses, we identified the prodrug-equivalent compound leporin C that is converted into leporin B by the action of the hCYP isoenzyme 3A4. By employing various prodrug-activating enzymes and microbes that biosynthesize diverse arrays of natural products, we should be able to probe wider biosynthetic space for identification of interesting prodrug-type natural products.