Inhibition of VEGF₁₆₅-Induced Angiogenesis by Gold Nanoparticles through HQGQH as the Primary Binding Site

Inhibition of vascular endothelial growth factor 165 (VEGF₁₆₅)-mediated angiogenesis is a promising anticancer strategy. While gold nanoparticles (AuNPs) are effective inhibitors of VEGF₁₆₅, the precise molecular interface of their interactions with VEGF₁₆₅ is unclear, hindering rational therapeutic design. This study combined molecular dynamics (MD) simulations and experimental techniques to map the binding interface between AuNPs and VEGF₁₆₅. We discovered that AuNPs preferentially bind to the HQGQH motif on VEGF₁₆₅. Functional assays showed that this binding inhibited VEGF₁₆₅-induced endothelial cell migration and angiogenesis. The necessity of the HQGQH motif was unequivocally demonstrated by mutagenesis. A 5A-substituted VEGF₁₆₅ mutant exhibited markedly reduced AuNP binding affinity across binding free energy calculations, ELISA, and ζ potential measurements, and its angiogenic activity became resistant to AuNP inhibition to some extent. These findings reveal the molecular basis of AuNP-mediated VEGF₁₆₅ blockade, opening avenues for designing targeted antiangiogenic nanomedicines.