新型氟核苷类似物NCC在肝细胞模型中抑制拉米夫定抗性乙型

StephenW

Braz J Infect Dis. 2018 Dec 23. pii: S1413-8670(18)30497-5. doi: 10.1016/j.bjid.2018.11.005. [Epub ahead of print]
Novel fluoronucleoside analog NCC inhibits lamivudine-resistant hepatitis B virus in a hepatocyte model.
Zhang J1, Wang Y2, Peng Y1, Qin C3, Liu Y1, Li J3, Jiang J1, Zhou Y4, Chang J5, Wang Q1.
Author information

1
    The First Affiliated Hospital of Zhengzhou University, Academy of Medical and Pharmaceutical Science, Henan Key Laboratory for Pharmacology of Liver Diseases, Zhengzhou Shi, China.
2
    Zhengzhou University, School of Pharmaceutical Sciences, Zhengzhou Shi, China.
3
    The First Affiliated Hospital of Zengzhou University, Department of Pharmacy, Zhengzhou Shi, China.
4
    The First Affiliated Hospital of Zengzhou University, Department of Pharmacy, Zhengzhou Shi, China. Electronic address: [email protected].
5
    College of Chemistry and Molecular Engineering Zhengzhou University, Zhengzhou Shi, China.

Abstract

Antiviral drug resistance is the most important factor contributing to treatment failure using nucleos(t)ide analogs such as lamivudine for chronic infection with hepatitis B virus (HBV). Development of a system supporting efficient replication of clinically resistant HBV strains is imperative, and new antiviral drugs are needed urgently to prevent selection of drug-resistant HBV mutants. A novel fluorinated cytidine analog, NCC (N-cyclopropyl-4'-azido-2'-deoxy-2'-fluoro-β-d-cytidine), was recently shown to strongly inhibit human HBV in vitro and in vivo. This study was designed to evaluate the antiviral activity of NCC against lamivudine-resistant HBV. We generated a stable cell line encoding the major pattern of lamivudine-resistant mutations rtL180M/M204V and designated it "HepG2.RL1". Immuno-transmission electron microscopic examination and enzyme-linked immunosorbent assay were used to detect secretion of HBV-specific particles and antigens. Quantification of extracellular DNA and intracellular DNA of HepG2.RL1 cells by quantitative real-time polymerase chain reaction revealed >625-fold and >5556-fold increases in the 50% inhibitory concentration of lamivudine, respectively, compared with that for the wild-type virus. The results showed that NCC inhibited DNA replication and HBeAg production in wild-type or lamivudine-resistant HBV in a dose-dependent manner. In conclusion, screening for antiviral compounds active against lamivudine-resistant HBV can be carried out with relative ease using hepG2.RL1 cells. NCC is a potential antiviral agent against wild-type HBV and clinical lamivudine-resistant HBV and deserves evaluation for the treatment of HBV infection.
KEYWORDS:

HepG2.RL1 cells; Hepatitis B virus; Lamivudine-resistant; NCC; rtL180M/M204V

PMID:
    30586543
DOI:
    10.1016/j.bjid.2018.11.005

StephenW

Braz J Infect Dis。 2018年12月23日.pii：S1413-8670（18）30497-5。 doi：10.1016 / j.bjid.2018.11.005。 [提前打印]
新型氟核苷类似物NCC在肝细胞模型中抑制拉米夫定抗性乙型肝炎病毒。
Zhang J1，Wang Y2，Peng Y1，Qin C3，Liu Y1，Li J3，Jiang J1，Zhou Y4，Chang J5，Wang Q1。
作者信息

1
    郑州大学第一附属医院，医学科学院，河南省肝病药理重点实验室，郑州市，中国。
2
    郑州大学药学院，郑州石，中国。
3
    中国郑州市增城大学第一附属医院药学系。
4
    中国郑州市增城大学第一附属医院药学系。电子地址：[email protected]。
五
    郑州大学化学与分子工程学院，郑州石，中国。

抽象

抗病毒药物耐药性是使用核苷（酸）类似物如拉米夫定治疗乙型肝炎病毒（HBV）慢性感染导致治疗失败的最重要因素。开发支持临床抗性HBV毒株的有效复制的系统是必要的，并且迫切需要新的抗病毒药物以防止选择耐药的HBV突变体。最近显示新的氟化胞苷类似物NCC（N-环丙基-4'-叠氮基-2'-脱氧-2'-氟-β-d-胞苷）在体外和体内强烈抑制人HBV。本研究旨在评估NCC对拉米夫定耐药HBV的抗病毒活性。我们产生了编码拉米夫定抗性突变rtL180M / M204V的主要模式的稳定细胞系，并将其命名为“HepG2.RL1”。免疫透射电子显微镜检查和酶联免疫吸附测定用于检测HBV特异性颗粒和抗原的分泌。通过定量实时聚合酶链反应定量分析HepG2.RL1细胞的细胞外DNA和细胞内DNA，分别显示拉米夫定50％抑制浓度> 625倍和> 5556倍，与野生型相比病毒。结果表明，NCC以剂量依赖的方式抑制野生型或拉米夫定耐药HBV中的DNA复制和HBeAg产生。总之，使用hepG2.RL1细胞可以相对容易地筛选对拉米夫定抗性HBV具有活性的抗病毒化合物。 NCC是一种抗野生型HBV和临床拉米夫定耐药性HBV的潜在抗病毒药物，值得对HBV感染的治疗进行评估。
关键词：

HepG2.RL1细胞;乙型肝炎病毒;拉米夫定抗性; NCC; rtL180M / M204V

结论：
    30586543
DOI：
    10.1016 / j.bjid.2018.11.005

StephenW

https://www.sciencedirect.com/sc ... 18304975?via%3Dihub