Design, Synthesis, and Biological Evaluation of APN and AKT Dual-Target Inhibitors

[Image: see text] Herein a novel series of APN and AKT dual inhibitors were derived from the clinical AKT inhibitor AZD5363. It was demonstrated that most compounds exhibited remarkable APN inhibitory activities with the most potent compound 8b (IC(50) = 0.05 ± 0.01 μM) being over 70-fold more potent than the approved APN inhibitor bestatin (IC(50) = 3.64 ± 0.56 μM). The moderate AKT inhibitory potencies of target compounds were also confirmed, with 5f and 5h possessing AKT1 IC(50) values of 0.12 and 0.27 μM, respectively. More importantly, the APN IC(50) values of 5f and 5h were 0.96 and 0.21 μM, respectively, indicating their balanced APN and AKT dual inhibition. HUVEC tube formation assays confirmed the superior APN inhibitory activities of 5f and 5h relative to bestatin at the cellular level. Western blot analysis demonstrated that 5h could effectively inhibit the phosphorylation of GSK3β, the intracellular substrate of AKT.