Inhibition of VEGF ₁₆₅ -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.