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In the present study, hydroxy-2-methyl-4H-pyran-4-one was isolated from the methanol root extract of Senecio laetus and was identified by FT-IR, ¹H, ¹³C NMR and GC-MS. The larvicidal potential of the compound 3-hydroxy-2-methyl-4H-pyran-4-one was evaluated against the 4^th instar larvae of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus at concentrations ranging from 20 to 1.25 ppm under laboratory conditions. The compound showed 100% mortality at 20 ppm against all the tested mosquitoe species and the LC₅₀ and LC₉₀ values were 1.22 and 7.25 ppm (An. stephensi), 2.10 and 99.84 ppm (Ae. aegypti) and 3.88 and 12.47 ppm (Cx. quinquefasciatus), respectively after 24 h of exposure period. In silico molecular docking study results reflects that hydroxy-2-methyl-4H-pyran-4-one compound showed highest binding affinity with OBP of Cx. quinquefasciatus (Glide energy score – 7 …