2-Phenylethynyl-Butyltellurium (PEBT) is a synthetic organotellurium compound that has shown various pharmacological properties on mammals without any signs of toxicity, but its effects on insects have not been reported before. Therefore, the aim of this study was to assess whether acute exposure to PEBT would promote an insecticidal effect against Drosophila melanogaster. The flies were exposed to three concentrations of PEBT (0.325 µmol L-1, 1.300 µmol L-1, and 5.200 µmol L-1) and a control solution (vehicle), using 450 flies per treatment (three repetitions of 150 flies), for 48 hours. Negative geotaxis and open field tests were performed (in vivo) after 24 and 48h, and acetylcholinesterase (AChE) activity was assessed (ex vivo) after 48h. Also, the mortality rate, 50% Lethal Concentration (LC50), 80% Lethal Concentration (LC80), and 95% Lethal Concentration (LC95) were calculated. Our results show that PEBT presented insecticidal activity against Drosophila melanogaster at all tested concentrations, which caused locomotor impairment and increased acetylcholinesterase activity in the flies' heads.