Chemical inhibitors of cholesterol binding function in a mosquito sterol carrier protein, AeSCP-2, were identified through high throughput screen of a small chemical library. AeSCP-2 inhibitors bind to purified recombinant AeSCP-2 with high affinities and showed inhibitory effects on cholesterol incorporation in cultured cells. AeSCP-2 inhibitors were tested in mosquito and tobacco hornworm larvae and showed potent insecticidal activities. The results indicate that a sterol carrier protein-2 has functional similarity in two different insect species. The discovery also suggests that targeting cholesterol metabolism for the development of new insecticides is highly plausible.