Glypican-3-targeted pH-responsive dual-drug co-loaded nanoliposomes for precision combined chemotherapy and anti-metastasis of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) chemotherapy is limited by poor drug selectivity, severe systemic toxicity, and frequent drug resistance. This study aimed to construct a novel pH-responsive nanoliposome system co-loaded with paclitaxel (PTX) and doxorubicin (ADR), functionalized with anti-glypican-3 (GPC3) antibodies, for precision combined chemotherapy and anti-metastasis of HCC.

Methods

PTX-loaded nanoliposomes were first prepared via reverse evaporation, followed by co-loading with ADR and bovine serum albumin to form PTX-ADR-BNP. GPC3 antibodies were conjugated onto the surface to obtain GPC3-PTX-ADR-BNP. The system was evaluated for particle size, zeta potential, encapsulation efficiency, and pH-responsive release. Cytotoxicity was assessed against HCC and normal cells. In vivo antitumor efficacy, biosafety, and anti-metastatic effects were evaluated using subcutaneous xenograft and lung metastasis models.

Results

GPC3-PTX-ADR-BNP exhibited uniform particle size (~180 nm), high encapsulation efficiencies (>96% for PTX, >95% for ADR), and pH-responsive release (~80% at pH 5.3 within 16 h, slow release at pH 7.4). In vitro, it showed potent cytotoxicity against MHCC97H and HuH-7 cells (half-maximal inhibitory concentration: 41.21 and 54.19 μg/mL) with minimal toxicity to normal cells. In vivo, the system prolonged plasma half-life, enhanced tumor accumulation, and achieved >59% tumor inhibition. Biosafety assessments, including hemolysis (<1% at all tested concentrations), serum biochemistry, hematological parameters, and histopathological examination of major organs, showed no significant toxicity compared to the control group. In the pulmonary metastasis model, it effectively suppressed lung metastasis, outperforming free drug and non-targeted groups.&#xD;

Conclusion

GPC3-PTX-ADR-BNP integrates GPC3-mediated active targeting with pH-responsive release for efficient anti-tumor and anti-metastatic effects, with high biocompatibility and low toxicity, offering a promising strategy for precision HCC therapy.