HetCA/STOCSY-Guided Dereplication of Artemisia arborescens Reveals Furofuran Lignans with Antileishmanial Effects.

Natural products represent an important source of bioactive compounds; however, their complexity often challenges conventional discovery approaches. In this study, an ethyl acetate extract of Artemisia arborescens (Vaill.) L. was investigated prior to any isolation using an integrated statistical spectroscopy-driven dereplication strategy combining the HeteroCovariance Approach (HetCA), Statistical Total Correlation Spectroscopy (STOCSY), 1H NMR, and LC-HRMS/MS. HetCA illustrated candidate features correlated with antileishmanial activity, predominantly associated with furofuran lignans. STOCSY facilitated metabolite annotation and distinction of structurally similar metabolites, enabling the identification of a novel lignan and a previously unreported compound in A. arborescens. Targeted isolation verified STOCSY-guided annotations, while biological evaluation of the pure metabolites revealed that compounds 1-3 exhibit moderate activity against Leishmania promastigotes, despite originating from a crude extract exhibiting negligible antileishmanial activity under the initial screening conditions. These findings demonstrate that HetCA can reveal bioactive compounds independently of extract-level activity and highlight its potential as a robust tool for broadening natural product drug discovery beyond conventional bioactivity-guided approaches.