Abstract: Background and Objective: Brusatol is one of quassinoid, derived from Makassar fruit (Brucea javanica (L.) Merr.) and belong to the family of Simaroubaceae. Brusatol has been proven to have insecticidal and anti-feeding activity on larvae of Spodoptera exigua. This study aimed to investigate the larvicidal activity and behavioral response to the larvae of A. aegypti after administration of brusatol and to demonstrate cytotoxicity activity in vero cells. Materials and Methods: Larvicidal activity was monitored using bioassay test. The behavioral response of larval A. aegypti was observed after treatment with various brusatol concentrations and compared directly with control every 10, 30 min and 1, 5 and 24 h. The examination of brusatol cytotoxicity on vero cells performed by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide) method. Results: The results of this study showed that the behavioral response of A. aegypti larvae after treatment with brusatol has similarity with the behavioral response after treatment with temephos 1 ppm. The behavioral response of A. aegypti larvae showed symptoms of a nerve poison that is excitation, convulsions, tremors and paralysis. The lethal concentration of 50 and 90 (LC50, LC90) to A. aegypti larvae were 0.669±0.106 and 8.331±0.060 ppm, respectively. Brusatol cytotoxic activity on vero cells is low, with IC50 values worth 1402.029±0.164 mg mL–1. Conclusion: The results of this study proved the potential of brusatol as biolarvicide to control the mosquito vector with low levels of cytotoxicity.