Inverse in silico-in vitro fishing of unexpected paroxetine kinase targets from tumor druggable kinome.

The serotonin selective reuptake inhibitor paroxetine has been clinically observed to reposition a significant suppressing potency on human tumors by unexpectedly targeting diverse kinase pathways involved in tumorigenesis. Here, we describe an inverse in silico-in vitro strategy to fish potential kinase targets using the paroxetine as bait. This is different (inverse) to the traditional drug discovery process that commonly screens small-molecule inhibitors for a specific kinase target. In the procedure, cell viability assays demonstrate that paroxetine has strong cytotoxicity on human tumor cell lines. Various protooncogene protein kinases are ontologically/manually enriched to define a druggable kinome, and a systematic interaction profile of paroxetine with the kinome is created, which indicates that paroxetine can potentially bind to some known targets or key regulators of human tumors. Kinase assays determine that paroxetine can effectively inhibit c-Src family kinases at nanomolar or micromolar levels. It is observed that the paroxetine ligand forms a tightly packed interface against the active site of these unexpected kinase targets to constitute several specific hydrogen bonds/π-π/cation-π stackings and a number of nonspecific hydrophobic/vdW contacts, while exposing a portion of molecular surface to solvent. More significantly, the ligand adopts two distinct binding modes (i.e., class I and class II) to interact with different kinases; the class-I mode has a higher stability and inhibitory activity than class-II mode. Steric clash seems to cause the ligand flipping from class I to class II. Graphical abstract.