The growing resistance of Aedes aegypti (L.) to conventional insecticides presents a major challenge in arbovirus control, necessitating the exploration of alternative insecticidal chemistries. Spiromesifen, derived from spirocyclic tetronic acids, is widely used against agricultural pests and is crucial in resistance management due to its unique lipid synthesis inhibition. This study evaluates the insecticidal activity of spiromesifen against temephos-resistant Ae. aegypti populations, focusing on larval body weight, volume, biochemical composition, and adult female reproductive potential. Spiromesifen demonstrated effective larvicidal activity, significantly reducing adult emergence. Resistance to spiromesifen was not observed, with resistance ratios (RR50, RR90) ranging from 0.36- to 3.31-fold. Larvae exposed to LC50 showed significant reductions in body weight and volume, and reduced carbohydrate, lipid, and protein contents. Enhanced catalase activity and malondialdehyde levels indicated increased oxidative stress and lipid peroxidation, highlighting its effects on lipid metabolism. Spiromesifen also exhibited sterilizing effects, significantly reducing fecundity and fertility in adult females, thereby impacting Ae. aegypti reproductive capacity. These findings highlight the potential of spiromesifen as a component of integrated vector management strategies, especially in regions with prevalent insecticide resistance in Ae. aegypti, serving as an effective larvicide and impacting adult reproductive outcomes.