A Combined Approach for Rapid Dereplication of Herb–Drug Interaction Causative Agents in Botanical Extracts─A Molecular Networking Strategy To Identify Potential Pregnane X Receptor (PXR) Modulators in Yohimbe

Despite promising preliminary biology, natural products isolation efforts may be confounded when the active compound is not isolated during bioassay-guided purification or classical pharmacognostic research investigations. A more rational isolation procedure connecting the polypharmacology of an herb to its individual constituents must be applied to better detect bioactive molecules before tedious analytical steps are considered. While <i>Pausinystalia johimbe</i> (yohimbe) has been traditionally used in herbal medicine as a general tonic, an aphrodisiac, a performance enhancer, and an integral part of various dietary supplements, the hydroethanolic extract of yohimbe was identified to possess at least 3-4-fold induction of the pregnane X receptor (PXR) at 30 μg/mL, a key nuclear receptor implicated in adverse interactions, <i>viz</i>., herb-drug interactions (HDIs). For rapid dereplication of potential HDI agents within yohimbe, a novel MS/MS-based molecular networking analysis was integrated with <i>in vitro</i> data and <i>in silico</i> analysis of activity at PXR. Analysis of the molecular network of biologically active fractions resulted in the dereplication of three oxindole alkaloids, 14 indole alkaloids, and eight <i>N</i>-oxide alkaloids as the primary causative agents for PXR induction. The findings of this study indicate that this strategy could effectively guide the rapid dereplication of bioactive causative agents within complex botanical extracts. Additionally, it serves as a proof-of-concept for using integrated MS/MS-based molecular networking analysis to assess the safety profile of botanical supplements.