对乙型肝炎病毒核心蛋白中天然存在的T109 / V124突变的GLS4（HA

StephenW

J Med Virol. 2017 Apr 12. doi: 10.1002/jmv.24830. [Epub ahead of print]
Influences on viral replication and sensitivity to GLS4, a HAP compound, of naturally occurring T109/V124 mutations in hepatitis B virus core protein.
Wang J1, Zhang H1, Zhang Y2, Jiang D3, Li J2, Goldmann S2, Ren Q2, Fei R1, Wang X1, Wei L1.
Author information

1    Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Peking University, Beijing 100044, China.
2    State Key Laboratory of Anti-Infective Drug Development, Sunshine Lake Pharma Co.,Ltd, Dong Guan, 523808, China.
3    Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, 100015, China.

Abstract

Heteroaryldihydropyrimidine (HAP) compounds inhibit HBV replication by binding to a hydrophobic pocket at the interface between hepatitis B virus core protein (HBcAg) dimer, which interrupts capsid assembly by changing the kinetics and thermodynamics during this process. Structure biological studies have identified several amino acids in HBcAg crucial for compound binding. Here we investigated the polymorphisms of T109 and V124 amino acids in HBV sequences submitted to GenBank database. Naturally occurring T109 and V124 and/or possible compensatory mutations in neighbored amino acids were introduced into HBV-expressing plasmids. Viral replication competence and sensitivity to GLS4, a HAP compound, were evaluated using transient transfection and in vitro infection cell models. All tested mutations in these amino acids led to decreasing viral DNA replication at different levels. Specially, T109N and all V124 mutants caused severe deficiencies in viral plus-strand DNA synthesis. T109I single mutation and all T109S/M/C/N mutations impaired HBeAg secretion. T109I showed modestly decreased sensitivities with IC50 3.3 to 6.8 folds higher than wild-type virus. In vitro infection assay showed T109N and all V124 mutants failed to synthesize cccDNA and following viral proteins. The other mutants, however, produced functional cccDNA pools as wild-type virus did. Taken together, we profiled the competences of viral replication and sensitivities to capsid inhibitor of naturally existing mutations in T109 and V124. This will help to understand the possible antiviral resistance issues in future clinical applications of capsid inhibitors. This article is protected by copyright. All rights reserved.

This article is protected by copyright. All rights reserved.
KEYWORDS:

Capsid inhibitor; Hepatitis B virus; Heteroaryldihydropyrimidine; Mutation

PMID:
    28401569
DOI:
    10.1002/jmv.24830

StephenW

J Med Virol 2017年4月12日。doi：10.1002 / jmv.24830。 [提前印刷]
对乙型肝炎病毒核心蛋白中天然存在的T109 / V124突变的GLS4（HAP化合物）的病毒复制和敏感性的影响。
王J 1张张H1张Y2 Jiang Jiang Jiang Jiang Li Li Li，Ren Ren Ren Ren i i i i Wei Wei Wei Wei Wei Wei Wei Wei Wei Wei Wei
作者信息

1
    北京大学人民医院北京大学肝脏病研究所，北京市北京大学丙型肝炎病毒免疫治疗重点实验室，北京100044。
2
    阳光湖药业有限公司抗感染药物开发国家重点实验室，东莞523808。
3
    首都医科大学北京Ditan医院传染病研究所，北京100015。

抽象

杂芳基二氢嘧啶（HAP）化合物通过结合乙型肝炎病毒核心蛋白（HBcAg）二聚体之间的界面处的疏水口袋来抑制HBV复制，其通过在该过程中改变动力学和热力学来中断衣壳组装。结构生物学研究已经确定了HBcAg中的几种氨基酸对复合结合至关重要。在这里我们调查了提交给GenBank数据库的HBV序列中T109和V124氨基酸的多态性。将自然发生的T109和V124和/或相邻氨基酸中的可能的补偿性突变引入HBV表达质粒。使用瞬时转染和体外感染细胞模型评价了对GLS4（一种HAP化合物）的病毒复制能力和敏感性。这些氨基酸中的所有测试突变导致不同水平降低病毒DNA复制。特别地，T109N和所有V124突变体引起病毒加上链DNA合成的严重缺陷。 T109I单突变和全部T109S / M / C / N突变均使HBeAg分泌受损。 T109I显示出适度降低的敏感性，IC50比野生型病毒高3.3〜6.8倍。体外感染测定显示T109N和所有V124突变体未能合成cccDNA和以下病毒蛋白。然而，其他突变体产生作为野生型病毒的功能性cccDNA库。总而言之，我们分析了病毒复制和敏感性对T109和V124中天然存在的突变的衣壳抑制剂的能力。这将有助于了解在未来的衣壳抑制剂临床应用中可能的抗病毒药物抗性问题。本文受版权保护。版权所有。

本文受版权保护。版权所有。
关键词：

衣壳抑制剂;乙型肝炎病毒杂芳基二氢嘧啶;突变

PMID：
    28401569
DOI：
    10.1002 / jmv.24830

newchinabok

核衣壳抑制剂有抗药性吗？

StephenW

回复 newchinabok 的帖子

以上研究表明GLS4也是有效对抗(核衣壳抑制)T109和V124突变型的HBV病毒.

阳光湖药业应公布GLS4临床试验II的结果.

newchinabok

回复 StephenW 的帖子

感谢答疑解惑，核衣壳，核心蛋白抑制剂是离成功最近的药，很关注。今年下半年几个药就见分晓。期待中。莫非赛定，虽然不成功，但意义重大。