论文标题
sofosbuvir封装的壳聚糖纳米颗粒的细胞毒性和抗HCV-4特性的体外评估
In Vitro Evaluation of Cytotoxic and Anti-HCV-4 Properties of Sofosbuvir Encapsulated Chitosan Nanoparticles
论文作者
论文摘要
Sofosbuvir是一种有效的HCV NS5B核苷酸聚合酶抑制剂，具有广泛的基因型覆盖范围，并且耐药性较低。虽然临床研究提供了沙发布韦对丙型肝炎病毒基因型4（HCV-4）的治疗的有效性，但是许多副作用均已报道。为了减少这些副作用并改善药物的抗病毒活性，将sofosbuvir封装在壳聚糖纳米颗粒（CNP）中，以产生sofosbuvir封装的壳聚糖纳米颗粒（SCNPS）。对Sofosbuvir和SCNP进行了3D分子模拟和动力学，以评估HCV-4 NS3蛋白酶，NS5B聚合酶和HCV解旋酶相对于它们的催化活性和药物敏感性的活性位点。最终在肝母细胞瘤细胞（HUH7）上评估了产生的SCNP，以提高其抗病毒效率。 结果：FTIR证实了SCNP的建议的化学结构。 SCNP的平均粒径和表面电荷分别为137 +/- 34 nm和29 +/- 9.6 mV。封装效率为80％，负载效率为6％。 SCNP与HCV -4 NS3，NS5B和NS5A的结合亲和力分别为-156.512，-154.603和-131。但对于Sofosbuvir，基于Moldock评分，分别为-127.581，-131.535和-167。在48小时的细胞暴露后，使用高达100μM的SCNP的HCV-4感染HuH7细胞均未显示出明显的细胞毒性和遗传毒性作用。最后，与未处理的细胞相比，在24小时暴露于100μMSCNP后，观察到HCV RNA的完全消失。
Sofosbuvir is a potent HCV NS5B nucleotide polymerase inhibitor with broad genotypic coverage and low risk of developing drug resistance. While clinical studies have provided the effectiveness of sofosbuvir for treatment of patients with hepatitis C virus genotype 4 (HCV-4), however, many side-effects were reported. To reduce those side effects and improve the antiviral activity of the drug, sofosbuvir was encapsulated into chitosan nanoparticles (CNPs) to produce sofosbuvir encapsulated chitosan nanoparticles (SCNPs). 3D Molecular simulation and dynamics were made for sofosbuvir and SCNPs to evaluate the active sites of HCV-4 NS3 protease, NS5B polymerase and HCV helicase relative to both their catalytic activities and drug susceptibilities. The produced SCNPs were finally evaluated on hepatoblastoma cells (Huh7) for their antiviral efficiency. Results: The suggested chemical structure for SCNPs was confirmed by FTIR. The average particle size and surface charge of SCNPs were 137 +/- 34 nm and 29 +/- 9.6 mV respectively. The Encapsulation efficiency was 80% and the loading efficiency was 6%. The binding affinity of SCNPs with HCV-4 NS3, NS5B and NS5A were -156.512, -154.603 and -131 respectively. But for sofosbuvir were -127.581, -131.535 and -167 respectively based on the MolDock score. The treatment of HCV- 4 infected Huh7 cells with up to 100 μM of SCNPs neither showed significant cytotoxic nor genotoxic effects after 48 hours of cell exposure. Finally, a complete disappearance of HCV RNA was seen after 24 hours of exposure to 100 μM of SCNPs when compared with the untreated cells.
