Novel Allosteric Inhibitors of Deoxyhypusine Synthase Against Malignant Melanoma: Design, Synthesis, and Biological Evaluation

Based on the novel allosteric site of deoxyhypusine synthase (DHPS), two series of 30 novel 5-(2-methoxyphenoxy)-2-phenylpyrimidin-4-amine derivatives as DHPS inhibitors were designed and synthesized. Among them, compound , with the best DHPS inhibitory potency (IC = 0.014 μM), exhibited excellent inhibition against melanoma cells, which was superior to that of GC7. Besides, molecular docking and molecular dynamics (MD) simulations further proved that compound was tightly bound to the allosteric site of DHPS. Flow cytometric analysis and enzyme-linked immunosorbent assay (ELISA) showed that compound could inhibit the intracellular reactive oxygen species (ROS) level. Furthermore, by western blot analysis, compound effectively activated caspase 3 and decreased the expressions of GP-100, tyrosinase, eIF5A2, MMP2, and MMP9. Moreover, both Transwell analysis and wound healing analysis showed that compound could inhibit the invasion and migration of melanoma cells. In the study, the tumor xenograft model showed that compound effectively inhibited melanoma development with low toxicity.