Exploring 6-Hydroxy-3-Aryl/Heteroarylcoumarins as Promising Candidates Against Trypanosoma cruzi.

∼ 10 μM). Mechanistic analysis using a washout assay revealed that, rather than eliminating intracellular forms, 1f prevents the release of new trypomastigotes. Our findings suggest that 1f targets the amastigote-to-trypomastigote differentiation pathway. The retention of a high intracellular parasitic load and the absence of trypomastigote release during the washout period strongly suggest that 1f acts as a selective inhibitor of metacyclogenesis. This insight positions the coumarin scaffold bearing a 2,3-dihydrobenzo[b][1,4]dioxine substituent at position 3 as a promising starting point for developing improved multi-stage therapeutic candidates against Trypanosoma cruzi.