Besides focusing urokinase (uPA) proteolytic activity on the cell membrane, the urokinase receptor (uPAR) is able to bind vitronectin (VN), via a direct binding site. Furthermore, uPAR interacts with other cell surface receptors, such as integrins, receptor tyrosin kinases and chemotaxis receptors, triggering cell-signalling pathways that promote tumor progression.(1) The ability of uPAR to coordinate binding and degradation of extracellular matrix and cell signalling makes it an attractive therapeutic target in cancer. We used structure-based virtual screening (SB-VS) (2) to search for small molecules targeting the uPAR binding site for VN. 41 compounds were identified and tested on uPAR-negative HEK-293 epithelial cells transfected with uPAR (uPAR-293 cells), using the parental cell line transfected with the empty vector (V-293 cells), as a control. Compounds, 6 and 37 selectively inhibited uPAR-293 cell adhesion to VN and the resulting changes in cell morphology and signal transduction, without exerting any effect on V-293 cells. 6 and 37 inhibited uPAR-293 cell binding to VN with IC50 values of 3.6 and 1.2 μM, respectively. Compounds 6 and 37 targeted S88 and R91, key residues for uPAR binding to VN but also for uPAR interaction with the f-MLF family of chemotaxis receptors (fMLF-Rs). As a consequence, 6 and 37 impaired uPAR-293 cell migration toward FCS, uPA and f-MLF, likely by inhibiting the interaction between uPAR and FPR1, the high affinity fMLF-R. Both compounds blocked in vitro extracellular-matrix invasion of several cancer cell types and could represent new promising leads for pharmaceuticals in cancer.(3)
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