Department of Pharmaceutics, College of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning, PR China.
Department of Science and Biotechnology, College of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning, PR China.
Int J Pharm. 2017 Mar 30;520(1-2):98-110. doi: 10.1016/j.ijpharm.2017.02.010. Epub 2017 Feb 4.
In order to overcome the shortcomings associated with the clinical application of norcantharidin (NCTD), including intense irritation and a short half-life, and to obtain a hepatocyte-selective liposome system with high encapsulation efficiency (EE) and low leakage, we synthesized a C alkyl chain norcantharimide derivative of NCTD (2-tetradecylhexahydro-1H-4,7-epoxyisoindole-1,3(2H)-dione, N-14NCTDA). Asialoglycoprotein receptor-targeted, galactosylated liposomes loaded with N-14NCTDA (GAL-Lipo) were prepared by the lipid film hydration method. GAL-Lipo with a satisfactory particle size of approximately 120nm has a higher encapsulation efficiency of more than 98.0%, which is markedly increased compared with NCTD loaded liposomes (EE%=47.6%). In addition, GAL-Lipo remained stable for at least 1 month at 4°C. In cytotoxicity assays, GAL-Lipo demonstrated stronger cytotoxicity effects (IC=24.58μmolL) on Hep G2 cells than free N-14NCTDA (100μmol/L) and conventional liposomes (Con-Lipo, 39.49μmol/L) without the GAL modification. GAL-Lipo can continuously accumulate in Hep G2 cells and be internalized into cells via two pathways, namely caveolin-dependent endocytosis and clathrin-dependent asialoglycoprotein receptors (ASGP-R) mediated endocytosis and produces considerably more significant cellular apoptosis. The results of vivo toxicity studies showed that GAL-Lipo dramatically reduced renal toxicity. In addition, GAL-Lipo has a markedly improved pharmacokinetic profile in vivo and a longer circulation time (AUC=6.700±2.964mgLh, t=1.347±0.519h) than Con-Lipo (AUC=2.319±0.121mgLh, t1/2z=0.413±0.238h). In conclusion, N-14NCTDA with an ideal logP is a better alternative for the treatment of primary hepatic carcinoma. GAL-Lipo offers an attractive strategy to specifically target hepatocytes via caveolin-dependent and clathrin-dependent asialoglycoprotein receptor-mediated endocytosis resulting in higher anticancer activity and fewer side-effects.
为了克服去甲斑蝥素（NCTD）在临床应用中的缺点，包括强烈的刺激性和半衰期短，并且获得具有高包封效率（EE）和低泄漏的肝靶向脂质体系统，我们合成了 NCTD 的 C 烷基链去甲斑蝥酰胺衍生物（2-十四烷基六氢-1H-4,7-环氧异吲哚-1,3(2H)-二酮，N-14NCTDA）。通过脂质体薄膜水化法制备了靶向去唾液酸糖蛋白受体的载有 N-14NCTDA 的半乳糖化脂质体（GAL-Lipo）。GAL-Lipo 的粒径约为 120nm，具有令人满意的包封效率，超过 98.0％，与载有 NCTD 的脂质体（EE％=47.6％）相比有显著提高。此外，GAL-Lipo 在 4°C 下至少稳定 1 个月。在细胞毒性测定中，与游离 N-14NCTDA（100μmol/L）和常规脂质体（Con-Lipo，39.49μmol/L）相比，GAL-Lipo 对 Hep G2 细胞具有更强的细胞毒性作用（IC=24.58μmol/L），而没有 GAL 修饰。GAL-Lipo 可以连续积聚在 Hep G2 细胞中，并通过两种途径被内化到细胞中，即小窝蛋白依赖性内吞作用和网格蛋白依赖性去唾液酸糖蛋白受体（ASGP-R）介导的内吞作用，并产生更显著的细胞凋亡。体内毒性研究结果表明，GAL-Lipo 显著降低了肾毒性。此外，GAL-Lipo 在体内具有明显改善的药代动力学特征和更长的循环时间（AUC=6.700±2.964mgLh，t=1.347±0.519h），优于 Con-Lipo（AUC=2.319±0.121mgLh，t1/2z=0.413±0.238h）。总之，具有理想 logP 的 N-14NCTDA 是治疗原发性肝癌的更好选择。GAL-Lipo 提供了一种有吸引力的策略，通过小窝蛋白依赖性和网格蛋白依赖性去唾液酸糖蛋白受体介导的内吞作用特异性靶向肝细胞，从而具有更高的抗癌活性和更少的副作用。
