Computational Predictive and Electrochemical Detection of Metabolites (CP-EDM) of Piperine

In this article we introduce a proof of concept strategy: Computational Predictive and Electrochemical Detection of Metabolites (CP-EDM) to expedite the discovery of drug metabolites. The use of a bioactive natural product, piperine, that has a well curated metabolite profile but has an unpredictable computational metabolism (Biotransformer v3.0) was selected. We developed an electrochemical reaction to oxidise piperine into a range of metabolites, which were detected by LC-MS. In turn, a series of chemically plausible metabolites were predicted based on ion-fragmentation patterns. These metabolites were docked into the active site of CYP3A4 using Autodock4.2 From the clustered low-energy profile of piperine in the active site it can be inferred that the most likely metabolic position of piperine (based on intermolecular distances to the Fe-oxo active site) is the benzo[d][1,3]dioxole motif. The metabolic profile was confirmed by literature comparison and the electrochemical reaction delivered plausible metabolites vide infra. Thus, demonstrating the power of the hyphenated technique of tandem electrochemical detection and computational evaluation of binding poses. Taken together, we outline a novel approach where diverse data sources are combined to predict and confirm a metabolic outcome for a bioactive structure.