Reverse translational approach to clarify the strong potency of enavogliflozin, a novel sodium-glucose cotransporter 2 inhibitor.

Enavogliflozin is a novel sodium-glucose cotransporter (SGLT2) inhibitor approved by the Korea Ministry of Food and Drug Safety for the treatment of type 2 diabetes. Clinical trials have demonstrated that enavogliflozin provides superior blood glucose-lowering effects and urinary glucose excretion (UGE) at a low dose of 0.3 mg compared with 10 mg of dapagliflozin. This study aimed to elucidate the structural characteristics of enavogliflozin and its major metabolites, M1 and M2, using a reverse translational approach and to investigate the mechanisms underlying its potent efficacy at approximately 30-fold lower doses than other SGLT2 inhibitors using an in vitro SGLT2 inhibition system. Molecular docking simulations revealed distinct binding modes for enavogliflozin and its metabolites relative to dapagliflozin and empagliflozin. The rigid conformation of enavogliflozin, conferred by its dihydrobenzofuran moiety, enforces hydrophobic and π-π stacking interactions, enhancing binding affinity, ligand-protein stability, and SGLT2 inhibition, suggesting a slower dissociation rate. Additionally, M1 and M2 exhibited inhibitory effects with SGLT1/2 dual inhibition and SGLT2 inhibition, respectively, indicating that both are pharmacologically active metabolites. Consequently, M1 and M2 significantly reduced blood glucose levels and increased UGE in rats. In conclusion, the clinical efficacy of enavogliflozin, including its potent glycemic-lowering effects and long-lasting UGE, is attributable to its structural features that promote hydrogen bonding and hydrophobic interactions with the galactose-binding motif, prolonged binding affinity to SGLT2, and the additional benefits of active metabolites M1 and M2. SIGNIFICANCE STATEMENT: The clinical efficacy of enavogliflozin is attributable to its high and sustained binding affinity for sodium-glucose cotransporter 2 (SGLT2) and the additional benefits conferred by active metabolites M1 and M2. The dihydrobenzofuran moiety of enavogliflozin is key to its distinctive inhibitory profile, contributing to its rigid conformation and optimal spatial orientation for effective π-π stacking interactions with Phe98, thereby enhancing its inhibitory potency. This structural feature distinguishes enavogliflozin from other SGLT2 inhibitors such as dapagliflozin and empagliflozin.