School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China.
The Second Hospital of Hunan University of Chinese Medicine, Changsha 410005, China.
Molecules. 2022 Oct 11;27(20):6786. doi: 10.3390/molecules27206786.
Cantharidin (CTD) is the major component of anticancer drugs obtained from and has a good inhibitory effect on several cancers, including hepatocellular carcinoma (HCC) and breast cancer. However, due to its toxicity, oral administration can cause various adverse reactions, limiting its clinical application. The aim of this work was to design glycyrrhetinic acid (GA)- and/or folate (FA)-modified solid lipid nanoparticles (SLNs) for the encapsulation of CTD to target HCC. Four CTD-loaded SLNs (cantharidin solid lipid nanoparticles (CSLNs), glycyrrhetinic acid-modified cantharidin solid lipid nanoparticles (GA-CSLNs), folate-modified cantharidin solid lipid nanoparticles (FA-CSLNs), and glycyrrhetinic acid and folate-modified cantharidin solid lipid nanoparticles (GA-FA-CSLNs)) were prepared by the emulsion ultrasonic dispersion method, and their physicochemical parameters were determined (particle size and distribution, morphology, zeta-potential, entrapment efficiency, drug loading, and hemolysis). Additionally, the antitumor activities of the four SLNs were evaluated comprehensively by tests for cytotoxicity, cell migration, cell cycle, apoptosis, cellular uptake, competition suppression assay, and in vivo tumor suppression assay. Four SLNs showed spherical shapes and mean diameters in the range of 75-110 nm with size dispersion (PDI) within the range of 0.19-0.50 and zeta-potential approximately -10 mV. The entrapment efficiency of CTD in SLNs was higher than 95% for all tested formulations, and no hemolysis was observed. Compared to GA-CSLNs or CSLNs, GA-FA-CSLNs and FA-CSLNs showed stronger cytotoxicity on hepatocellular carcinoma cells (HepG2), and the cytotoxicity of GA-FA-CSLNs on hepatocyte cells (L-02) was remarkably reduced compared with other formulations. GA-FA-CSLNs and FA-CSLNs also increased the inhibition of HepG2 cell migration, and FA-CSLNs had the highest apoptosis rate. The cell cycle results indicated that HepG2 cells were arrested mainly in the S phase and G2/M phase. Analysis of competition inhibition experiments showed that GA and FA ligands had targeted effects on HepG2 cells. The in vivo tumor inhibition experiment showed that GA-FA-CSLNs and FA-CSLNs had excellent tumor inhibition ability-their tumor inhibition rates were 96.46% and 89.92%, respectively. Our results indicate that GA-FA-CSLNs and FA-CSLNs have a promising future in the therapeutic intervention of HCC.
斑蝥素（CTD）是从 中提取的抗癌药物的主要成分，对包括肝癌（HCC）和乳腺癌在内的多种癌症具有良好的抑制作用。然而，由于其毒性，口服给药会引起各种不良反应，限制了其临床应用。本工作旨在设计甘草次酸（GA）和/或叶酸（FA）修饰的固体脂质纳米粒（SLNs）以封装 CTD 靶向 HCC。通过乳液超声分散法制备了四种载有 CTD 的 SLNs（斑蝥素固体脂质纳米粒（CSLNs）、甘草次酸修饰的斑蝥素固体脂质纳米粒（GA-CSLNs）、叶酸修饰的斑蝥素固体脂质纳米粒（FA-CSLNs）和甘草次酸和叶酸修饰的斑蝥素固体脂质纳米粒（GA-FA-CSLNs）），并测定了其理化参数（粒径和分布、形态、Zeta 电位、包封效率、载药量和溶血）。此外，通过细胞毒性、细胞迁移、细胞周期、细胞凋亡、细胞摄取、竞争抑制试验和体内肿瘤抑制试验综合评价了四种 SLNs 的抗肿瘤活性。四种 SLNs 均呈球形，粒径在 75-110nm 范围内，粒径分布（PDI）在 0.19-0.50 范围内，Zeta 电位约为-10mV。CTD 在 SLNs 中的包封效率均高于 95%，且无溶血。与 GA-CSLNs 或 CSLNs 相比，GA-FA-CSLNs 和 FA-CSLNs 对肝癌细胞（HepG2）的细胞毒性更强，GA-FA-CSLNs 对肝细胞（L-02）的细胞毒性明显低于其他制剂。GA-FA-CSLNs 和 FA-CSLNs 还增加了对 HepG2 细胞迁移的抑制作用，FA-CSLNs 的凋亡率最高。细胞周期结果表明 HepG2 细胞主要被阻滞在 S 期和 G2/M 期。竞争抑制实验分析表明，GA 和 FA 配体对 HepG2 细胞具有靶向作用。体内肿瘤抑制实验表明，GA-FA-CSLNs 和 FA-CSLNs 具有优异的肿瘤抑制能力-其肿瘤抑制率分别为 96.46%和 89.92%。我们的结果表明，GA-FA-CSLNs 和 FA-CSLNs 在 HCC 的治疗干预中具有广阔的应用前景。
