Repositioning of Small Molecules through the Inverse Virtual Screening in silico Tool: Case of Benzothiazole‐Based Inhibitors of Soluble Epoxide Hydrolase (sEH)

Computational techniques accelerate drug discovery byidentifying bioactive compounds for specific targets, optimizingmolecules with moderate activity, or facilitating the repositioning ofinactive items onto new targets. Among them, the Inverse VirtualScreening (IVS) approach is aimed at the evaluation of one or a smallset of molecules against a panel of targets for addressing targetidentification. In this work, a focused library of benzothiazole-basedcompounds was re-investigated by IVS. Four items, originallysynthesized and tested on bromodomain-containing protein 9 (BRD9)but yielding poor results, were critically re-analyzed, disclosing only apartial fit with 3D structure-based pharmacophore models, which, inthe meanwhile, were developed for this target. Afterwards, thesecompounds were re-evaluated through IVS on a panel of proteinsinvolved in inflammation and cancer, identifying soluble epoxidehydrolase (sEH) as a putative interacting target. Three items weresubsequently confirmed as able to inhibit sEH activity, spanning from70% up to 30% when tested at 10 μM. Finally, one benzothiazole-based compound emerged as the most promising inhibitor featuringan IC50 in the low micromolar range (IC50 = 6.62 ± 0.13 μM). Our dataconfirm IVS as a predictive tool for accelerating the targetidentification and repositioning processes