Virtual screening (VS) techniques are well-established tools in the modern drug discovery process. The availability knowledge of structural information, indicated by the increasing number of 3D protein structures and the readiness of free databases of commercially available small-molecules, supplies a broad platform for VS. This review summarizes current developments in virtual screening regarding chemical databases, highlighting achievements as well as challenges, with emphasis on a recent example of successful application for the identification of novel hit compounds for Trypanosoma cruzi sterol 14 α-demethylase (CYP51).