Harnessing Box-Behnken design to optimize erlotinib-niclosamide co-loaded liposomes, overcoming EGFR-mutated lung cancer resistance, and improving sensitivity via EGFR/STAT3 signaling pathway.

内容摘要

The co-administration of niclosamide (NCM) with erlotinib hydrochloride (ERL) has been shown to restore ERL sensitivity in resistant non-small cell lung cancer. However, the poor physicochemical properties of both molecules necessitate a specialized delivery system for improved delivery and efficacy. Box-Behnken design was used to evaluate the key variables affecting NCM-ERL-Liposome characteristics. The design yielded a broader design space for the selected factors that can achieve superior attributes, including particle size of 93.42 ± 2.41 nm, polydispersity index of 0.356 ± 0.01, zeta potential of -9.40 ± 0.59 mV, and maximal entrapment efficiencies (NCM: 77.97 ± 2.25%; ERL: 72.75 ± 1.49%). Trehalose showed better redispersion and good physical stability, as confirmed from ATR-IR/PXRD/DSC. The formulation demonstrated sustained dual-drug release, stability for three months at 4 ± 2 °C, and hemocompatibility with minimal hemolysis. In-vitro studies showed that NCM restored ERL sensitivity, with lower IC50 values (HCC827-P: 11.27 ± 4.04 µg/mL; HCC827-ER: 7.59 ± 0.62 µg/mL) than those of pure drugs or the physical mixture. The prepared NCM-ERL-Liposomes showed enhanced internalization, triggered caspase-3/7-mediated apoptosis (1.42-fold increase), and suppressed p-EGFR/p-STAT3, indicating adequate payload protection and targeted intracellular release. These results establish a translational platform that requires PK/PD studies in resistant NSCLC models to support precision oncology therapeutics. Article highlightsThis study discusses the challenge of chemotherapeutic resistance in NSCLC and explores dual-loaded liposomal nanocarriers as a potential solution.NCM-ERL-Liposomes were developed via thin-film hydration and systematic BBD optimization.Cryoprotectant-engineered glassy matrices mitigate lyophilization stress.The HCC827-ER model mimics clinical resistance (>6-fold increase in IC50).Maximal caspase-3/7 induction reverses resistance via apoptosis.Potent p-EGFR/p-STAT3 inhibition restores erlotinib sensitivity.