Molecular insights into the formation of polymeric nanoassemblies of the anticancer peptide PEN-FFW.

The peptide PEN-FFW has emerged as a candidate for hepatocellular carcinoma therapy, with selective efficacy and a favorable safety profile. Development of a suitable formulation has been hindered by its sensitivity to shear and limited physicochemical stability. In this work, we elucidated drug-excipient interactions and exploited them to design a stable nanoassembly. PEN-FFW was encapsulated within a carboxymethyl cellulose-poly(lactide-co-glycolide) core, stabilized by polysorbate 80. Binding and structural features were confirmed by quartz crystal microbalance with dissipation (QCM-D), Fourier transform infrared (FTIR), and dynamic light scattering (DLS), while molecular modeling suggested the formation of a pocket accommodating the peptide within the core. A four-factor, three-level Box-Behnken design was applied to optimize process parameters, yielding a 50 % increase in encapsulation efficiency and enhanced storage stability, as determined by desirability function analysis. The optimized formulation exhibited superior cellular uptake and cytotoxicity in vitro, and in vivo studies in mice demonstrated significantly greater hepatic retention relative to the free peptide.