Phytochemistry, Mode of Action Predictions, and Synergistic Potential of Hypenia irregularis Essential Oil Mixtures...

Aedes aegypti, also known as the yellow fever mosquito, presents a major public health challenge, highlighting the need for effective biorational agents for mosquito control. Here, we investigated the synergistic effects of essential oil mixtures derived from Hypenia irregularis that is a mint-family shrub native to Brazil's Cerrado biome, known as "alecrim do Cerrado", in combination with essential oils from noni (Morinda citrifolia), Brazilian mint ("salva-do-Marajó", Hyptis crenata), and lemongrass (Cymbopogon citratus) against Ae. aegypti. We conducted phytochemical analyses and assessed larvicidal, repellent, and oviposition deterrent activities. Using in silico methods, we predicted molecular interactions between key essential oil components and physiological targets involved in repellent action (odorant-binding protein AeagOBP1 and olfactory receptor Or31) and larvicidal activity (GABA and octopamine receptors, TRP channels, and acetylcholinesterase [AChE]). Major compounds identified included octanoic acid (23%; Hipe. irregularis × M. citrifolia), 2,5-dimethoxy-p-cymene (21.9%; Hipe. irregularis × Hypt. crenata), and citral (23.0%; Hipe. irregularis × C. citratus). Although individual oils showed strong larvicidal activity (Hipe. irregularis LC₅₀ = 2.35 µL/mL; Hypt. crenata = 2.37 µL/mL; M. citrifolia and C. citratus = 2.71 µL/mL), their mixtures did not display synergistic effects. Similarly, repellency and oviposition deterrence were comparable to DEET for individual oils but were not enhanced in mixtures. Notably, the Hipe. irregularis × C. citratus essential oil blend reduced oviposition deterrence. Molecular docking confirmed strong binding of major oil components to AeagOBP1 and Or31, supporting their role in repellency. For larvicidal effects, AChE showed the highest predicted binding affinity. Overall, our findings suggest that H. irregularis, Hypt. crenata, C. citratus, and M. citrifolia (alone or in 1:1 mixture) are promising, sustainable agents for A. aegypti control.