With the continuous evolution of insect resistance, it is a tremendous challenge to control the fall armyworm (Spodoptera frugiperda) with traditional insecticides.To solve this pending issue, a series of novel isoxazoline derivatives containing diaryl ether structures were designed and synthesized, and most of the target compounds exhibited excellent insecticidal activity.Based on the three-dimensional quant. structure-activity relationship (3D-QSAR) model anal., we further optimized the mol. structure with N-(4-((2-chloropyridin-3-yl)oxy)benzyl)-4-(5-(3,5-dichlorophenyl)-5-(trifluoromethyl)-4,5-dihydroisoxazol-3-yl)-2-methylbenzamide (L35) obtained and tested for its activity.Compound L35 (LC50 = 1.69 mg/L) exhibited excellent insecticidal activity against S. frugiperda, which was better than those of com. fipronil (LC50 = 70.78 mg/L) and indoxacarb (LC50 = 5.37 mg/L).The ELISA showed that L35 could upregulate the levels of GABA in insects.In addition, mol. docking and transcriptomic results also indicated that compound L35 may affect the nervous system of S. frugiperda by acting on GABA receptors.Notably, through high-performance liquid chromatog. (HPLC), we were able to obtain the two enantiomers of compound L35, and the insecticidal activity test revealed that S-(+)-L35 was 44 times more active than R-(-)-L35 against S. frugiperda.This study established the chem. basis and mechanistic foundations for the future development of pesticide candidates against fall armyworms.