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All structures were docked and scored using the Glide standard precision (SP) mode. The 10,000 top-ranked structures from the SP filter were redocked and rescored using the Glide extra precision (XP) mode. The complexes for the top-ranked 1,000 compounds resulting from the XP processing were submitted to further post-processing with the ChemBioServer. In order to filter out poses far from the energy minimum, we used the van der Waals filter of the server to remove compounds with steric clashes. The remaining compounds were then subjected to physicochemical property filtering based on the “Jorgensen rule of 3”  as well as toxicity filtering based on a database available in ChemBioServer, which contains known toxic moieties. Subsequently, a hierarchical clustering was performed for the remaining compounds using the Tanimoto coefficient and the Ward Clustering Linkage in order to maximize chemical diversity but at the same time minimize visual inspection efforts.
Finally, the resulting cluster representatives were visually inspected and the ten most promising compounds were purchased and submitted to in vitro assay testing. Four out of the ten purchased molecules inhibited the PI3Kα protein activity in vitro in μm concentrations, indicating that the workflow described herein can be successfully applied in drug discovery . “Minimum effective dose” is a medical term which refers to the amount of a substance you need for the results you want, and above which, the substance doesn’t increase in effectiveness.