U46 - Folic Acid-Modified Dual-Drug Nanocarriers to Overcome Drug Resistance in Hepatocellular Carcinoma

Introduction: : Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related deaths worldwide, with therapeutic resistance posing a significant challenge to effective treatment. Lenvatinib, a multi-kinase inhibitor, is used as a first-line treatment in unresectable HCC, however it frequently encounters resistance mediated by activation of the HGF/c-Met (Hepatocyte Growth Factor/Mesenchymal-epithelial transition factor) signaling axis. To overcome Lenvatinib resistance, we developed a folic acid-conjugated lipid-shell, poly lactic-co-glycolic acid (PLGA) polymer-core nanoparticle (NP) system to co-deliver Lenvatinib and a c-Met inhibitor, designed to suppress oncogenic signaling and restore drug sensitivity synergistically. Folic acid is a well-known targeting moiety that enables the NPs to bind with high affinity to folate receptors overexpressed in HCC. By leveraging this receptor-ligand interaction, folic acid-functionalized NPs can facilitate selective drug delivery to tumor cells.

Materials and

Methods: : Lenvatinib-encapsulated PLGA (Resomer, LA: GA, 50:50) NPs were prepared using a single emulsion solvent evaporation method. The folic acid was conjugated to DSPE-PEG-NH2 (1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-polyethylene glycol-amine) via a standard carbodiimide reaction to form DSPE-PEG-FA. The DSPE-PEG-FA containing the c-Met inhibitor was then used to coat the Lenvatinib-encapsulated PLGA NP core. NP size, surface charge, and morphology were characterized using dynamic light scattering (DLS), zeta potential analysis, and transmission electron microscopy (TEM), respectively. Drug loading efficiency and release kinetics were assessed via HPLC (High Performance Liquid Chromatography). Cellular uptake and cytotoxicity were evaluated in vitro using HepG2 hepatocellular carcinoma cell lines following stimulation using HGF (Hepatocyte growth factor).

Results, Conclusions, and Discussions:: Comprehensive physicochemical characterization confirmed uniform particle size (110.9 ±5.21 nm), negative surface charge (-20 ± 6.33 mV), and a Lenvatinib encapsulation efficiency of 40 ± 2.81% in the PLGA core. The conjugation efficiency of folic acid measured using UV-visible spectrometry was found to be around 89 ± 3.98 %. The NPs were cytocompatible and demonstrated dose-dependent uptake in HepG2 cells. The dual-drug loaded NPs exhibited markedly enhanced cytotoxicity in HGF-stimulated HepG2 cells in 5 days, than NPs containing Lenvatinib alone (Figure 1).

Conclusions: Altogether, these preliminary findings suggest that the dual-drug-loaded-NP formulation exhibited improved anti-cancer effects compared to that achievable using NPs containing a single drug, within 5 days of treatment. Ongoing and future studies are focused on the targeting and therapeutic effects of the NPs in in vitro and in vivo models of liver cancer.

Acknowledgements and/or References (Optional):: This work was supported by the National Institute of Health grant R37 CA283937 to J.U.M.