Formulation, interaction analysis, and invitro hepatocellular carcinoma studies of Rutin loaded lipid and polymeric nanoparticles

Hepatocellular carcinoma is a primary liver cancer with a high mortality rate worldwide. The limited efficacy and adverse effects of conventional chemotherapy have driven the exploration of novel therapeutic approaches, including Nano medicine-based anticancer agents. In this study, the best molecule was identified based on molecular-level interaction studies with the three anticancer pathway proteins. The best molecule was encapsulated with polymer and lipid to form nanoparticles and In vitro Hepatocellular carcinoma studies were performed to measure the activity. The interaction analysis revealed that Rutin binds effectively with all three proteins: folate receptor (5IZQ), vascular endothelial growth factor (5ABD), and CD44 (4PZ3), exhibiting binding energies of -44.5015 kcal/mol, -46.8331 kcal/mol, and -42.6949 kcal/mol, respectively. These results indicate that Rutin demonstrates stronger binding affinity compared to other anticancer agents by computation studies. In the formulation, Rutin-formed lipid nanoparticles exhibited higher encapsulation efficacy and drug-loading efficiency compared to polymeric nanoparticles. Dissolution studies of rutin-loaded lipid nanoparticles reveal a gradual release profile, reaching approximately 99% over 6 hours. This sustained release ensures extended drug activity within the tumor environment, improving therapeutic effectiveness. In vitro cytotoxicity studies demonstrated that Rutin-lipid nanoparticles had superior anticancer efficacy, with an IC50 value of 80.45 ± 0.05 µg/mL, compared to 120.45 ± 0.05 µg/mL for Rutin-polymeric nanoparticles against the Hep3B cell lines. Similarly, Fluorescence-based screening studies further confirmed the remarkable anti-cancer potential of Rutin-lipid nanoparticles, primarily by inducing apoptosis in Hep3B cells. These findings suggest that Rutin-lipid nanoparticles hold promise nano-formulations for targeting Hepatocellular carcinoma.