K-Ras is one of the most frequently mutated signal transducing human oncogenes. Ras signaling activity requires correct cellular localization of the GTPase. The spatial organization of K-Ras is controlled by the prenyl binding protein PDE未, which enhances Ras diffusion in the cytosol. Inhibition of the Ras鈥揚DE未 interaction by small molecules impairs Ras localization and signaling. Here we describe in detail the identification and structure guided development of Ras鈥揚DE未 inhibitors targeting the farnesyl binding pocket of PDE未 with nanomolar affinity. We report kinetic data that characterize the binding of the most potent small molecule ligands to PDE未 and prove their binding to endogenous PDE未 in cell lysates. The PDE未 inhibitors provide promising starting points for the establishment of new drug discovery programs aimed at cancers harboring oncogenic K-Ras.