Efficacy assessment of glycyrrhetinic acid-modified liposomes loaded with doxorubicin hydrochloride and cucurbitine B for synergistic treatment of hepatocellular carcinoma.

Objective: Hepatocellular carcinoma (HCC) is characterized by a high incidence rate, aggressive invasion and metastasis, and a significant postoperative recurrence rate. Targeted therapy plays a crucial role in the precise treatment of HCC. Studies have demonstrated that Glycyrrhetinic acid (GA) specific receptors are overexpressed on the surface of HCC cells. Doxorubicin hydrochloride (Dox), a widely used chemotherapy agent for anti-tumor treatment, but is associated with substantial toxic side effects. Cucurbitacin B (CuB) also demonstrates promising anti-tumor activity, but its poor water solubility and low bioavailability limit its clinical application. The combination of Dox and CuB can exert a synergistic effect, thereby enhancing the overall anti-tumor efficacy. Therefore, we have developed GA-modified liposomes loaded with Dox and CuB (GA-Dox/CuB-Lips) to achieve synergistic therapy for HCC.

Method: In this study, GA-Dox/CuB-Lips were prepared using the thin film dispersion method and ammonium sulfate gradient method. In vitro, we evaluated the cellular uptake and cytotoxicity of the liposomes, as well as their anti-tumor effects in inhibiting tumor proliferation, promoting tumor apoptosis, and suppressing invasion and metastasis. In vivo, the targeting properties of GA-Dox/CuB-Lips were assessed through in vivo imaging. A tumor growth curve was generated by establishing a heterotopic nude mouse model. Additionally, an in-situ HCC model was established and the anti-tumor effects of liposomes were evaluated using HE staining, histological analysis and immunofluorescence staining.

Results: We successfully prepared GA-Dox/CuB-Lips with a smooth, spherical morphology and uniform distribution. Both drugs exhibited high encapsulation efficiency, significantly enhancing the solubility of CuB. In vitro, GA-Dox/CuB-Lips demonstrated excellent targeting properties and exerted cytotoxic effects on Hepa1-6 cells, effectively inhibiting tumor cell proliferation, invasion, and metastasis while promoting tumor cell apoptosis. In vivo, GA-Dox/CuB-Lips selectively targeted tumor sites, disrupted tumor structures, inhibited tumor growth and proliferation, and promoted apoptosis.

Conclusion: GA-Dox/CuB-Lips exhibited excellent anti-HCC activity and represent a promising therapeutic approach for the treatment of HCC.