Diltiazem　and　glibenclamide　are　commonly　used　hypotensive　and　antidiabetic　drugs.　This　study　reports　the　discovery　of　the　potential　antitumor　and　antimetastatic　effects　of　these　two　drugs　using　a　structural　dynamics-driven　virtual　screening　targeting　urokinase　receptor　(uPAR).　Owing　to　uPAR’s　high　flexibility,　currently　resolved　crystal　structures　of　uPAR,　all　in　ligand-bound　states,　provide　limited　representations　of　its　physiological　conformation.　To　improve　the　accuracy　of　screening,　we　performed　a　long-timescale　molecular　dynamics　simulation　and　obtained　the　representative　conformations　of　apo-uPAR　as　the　targets　for　our　screening.　Experimentally,　we　demonstrated　that　diltiazem　and　glibenclamide　bound　uPAR　with　KD　values　in　the　micromolar　range.　In　addition,　both　compounds　effectively　suppressed　tumor　growth　and　metastasis　in　a　uPAR-dependent　manner　in　vitro　and　in　vivo.　This　work　not　only　provides　two　potent　uPAR　inhibitors　but　also　reports　a　proof-of-concept　study　on　the　potential　off-label　antitumor　and　antimetastatic　uses　of　diltiazem　and　glibenclamide.　©　2023　American　Chemical　Society.