Computational methods can be applied in drug development for the identification of novel lead candidates, but also for the prediction of pharmacokinetic properties and potential adverse effects, thereby aiding to prioritize and identify the most promising compounds. In principle, several techniques are available for this purpose, however, which one is the most suitable for a specific research objective still requires further investigation. Within this study, the performance of several programs, representing common virtual screening methods, was compared in a prospective manner. First, we selected top-ranked virtual screening hits from the three methods pharmacophore modeling, shape-based modeling, and docking. For comparison, these hits were then additionally predicted by external pharmacophore- and 2D similarity-based bioactivity profiling tools. Subsequently, the biological activities of the selected hits were assessed in vitro, which allowed for evaluating and comparing the prospective performance of the applied tools. Although all methods performed well, considerable differences were observed concerning hit rates, true positive and true negative hits, and hitlist composition. Our results suggest that a rational selection of the applied method represents a powerful strategy to maximize the success of a research project, tightly linked to its aims. We employed cyclooxygenase as application example, however, the focus of this study lied on highlighting the differences in the virtual screening tool performances and not in the identification of novel COX-inhibitors.

Department of Crop Sciences and Agroforestry Faculty of Tropical AgriSciences Czech University of Life Sciences Prague Kamycka 129 165 21 Prague 6 Suchdol Czech Republic

Institute of Pharmacy Pharmaceutical Chemistry and Center for Molecular Biosciences Innsbruck University of Innsbruck Innrain 80 82 6020 Innsbruck Austria

Laboratory of Plant Biotechnologies Institute of Experimental Botany AS CR v v i Rozvojova 263 165 02 Prague 6 Lysolaje Czech Republic

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