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Clinical use of phosphodiesterase-5 (PDE5) inhibitors is limited by several side effects due to weak isoform selectivity. Herein, a unique allosteric pocket of PDE5 is identified by molecular modeling and structural biology, which enables the discovery of highly selective PDE5 inhibitors from natural product evodiamine (EVO). The crystal structure of PDE5 with bound EVO derivative ( S )-7e revealed that binding of ( S )-7e to the novel allosteric pocket induced dramatic conformation changes in the H-loop with a maximum 24 Å movement of th