新型非杂芳基嘧啶（HAP）衣壳组装修饰剂在体外具有与HAP不

StephenW

Antiviral Res. 2018 Jul 19. pii: S0166-3542(18)30329-2. doi: 10.1016/j.antiviral.2018.07.011. [Epub ahead of print]
Novel non-heteroarylpyrimidine (HAP) capsid assembly modifiers have a different mode of action from HAPs in vitro.
Corcuera A1, Stolle K2, Hillmer S3, Seitz S4, Lee JY4, Bartenschlager R5, Birkmann A2, Urban A6.
Author information

1
    AiCuris Anti-infective Cures GmbH, Friedrich-Ebert-Str.475, 42117, Wuppertal, Germany; Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im Neuenheimer Feld 344, 69120, Heidelberg, Germany.
2
    AiCuris Anti-infective Cures GmbH, Friedrich-Ebert-Str.475, 42117, Wuppertal, Germany.
3
    Electron Microscopy Core Facility, Heidelberg University, Im Neuenheimer Feld 345, 69120, Heidelberg, Germany.
4
    Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im Neuenheimer Feld 344, 69120, Heidelberg, Germany.
5
    Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im Neuenheimer Feld 344, 69120, Heidelberg, Germany; German Center for Infection Research, Heidelberg Partner Site, Im Neuenheimer Feld 344, 69120, Heidelberg, Germany.
6
    AiCuris Anti-infective Cures GmbH, Friedrich-Ebert-Str.475, 42117, Wuppertal, Germany. Electronic address: [email protected].

Abstract

One of the most promising viral targets in current hepatitis B virus (HBV) drug development is the core protein due to its multiple roles in the viral life cycle. Here we investigated the differences in the mode of action and antiviral activity of representatives of six different capsid assembly modifier (CAM) scaffolds: three from the well-characterized scaffolds heteroarylpyrimidine (HAP), sulfamoylbenzamide (SBA), and phenylpropenamide (PPA), and three from novel scaffolds glyoxamide-pyrrolamide (GPA), pyrazolyl-thiazole (PT), and dibenzo-thiazepin-2-one (DBT). The target activity and antiviral efficacy of the different CAMs were tested in biochemical and cellular assays. Analytical size exclusion chromatography and transmission electron microscopy showed that only the HAP compound induced formation of aberrant non-capsid structures (class II mode of action), while the remaining CAMs did not affect capsid gross morphology (class I mode of action). Intracellular lysates from the HepAD38 cell line, inducibly replicating HBV, showed no reduction in the quantities of intracellular core protein or capsid after treatment with SBA, PPA, GPA, PT, or DBT compounds; however HAP-treatment led to a profound decrease in both. Additionally, immunofluorescence staining of compound-treated HepAD38 cells showed that all non-HAP CAMs led to a shift in the equilibrium of HBV core antigen (HBcAg) towards complete cytoplasmic staining, while the HAP induced accumulation of HBcAg aggregates in the nucleus. Our study demonstrates that the novel scaffolds GPA, PT, and DBT exhibit class I modes of action, alike SBA and PPA, whereas HAP remains the only scaffold belonging to class II inhibitors.
KEYWORDS:

Capsid; Capsid assembly modifier; Core protein; HBV; Heteroarylpyrimidine

PMID:
    30031759
DOI:
    10.1016/j.antiviral.2018.07.011

StephenW

抗病毒药物2018年7月19日.pii：S0166-3542（18）30329-2。 doi：10.1016 / j.antiviral.2018.07.011。 [提前打印]
新型非杂芳基嘧啶（HAP）衣壳组装修饰剂在体外具有与HAP不同的作用模式。
Corcuera A1，Stolle K2，Hillmer S3，Seitz S4，Lee JY4，Bartenschlager R5，Birkmann A2，Urban A6。
作者信息

1
    AiCuris Anti-infective Cures GmbH，Friedrich-Ebert-Str.475,42117，Wuppertal，Germany;海德堡大学传染病学系，分子病毒学，Im Neuenheimer Feld 344,69120，Heidelberg，Germany。
2
    AiCuris Anti-infective Cures GmbH，Friedrich-Ebert-Str.475,42117，Wuppertal，Germany。
3
    电子显微镜核心设施，海德堡大学，Im Neuenheimer Feld 345,69120，Heidelberg，Germany。
4
    海德堡大学传染病学系，分子病毒学，Im Neuenheimer Feld 344,69120，Heidelberg，Germany。
五
    海德堡大学传染病学系，分子病毒学，Im Neuenheimer Feld 344,69120，Heidelberg，Germany;德国感染研究中心，海德堡合作伙伴网站，Im Neuenheimer Feld 344,69120，Heidelberg，Germany。
6
    AiCuris Anti-infective Cures GmbH，Friedrich-Ebert-Str.475,42117，Wuppertal，Germany。电子地址：[email protected]。

抽象

当前乙型肝炎病毒（HBV）药物开发中最有希望的病毒靶标之一是核心蛋白质，因为它在病毒生命周期中具有多重作用。在这里，我们研究了六种不同衣壳组装修饰剂（CAM）支架代表的作用方式和抗病毒活性的差异：三种来自充分表征的支架杂芳基嘧啶（HAP），氨磺酰苯甲酰胺（SBA）和苯丙烯酰胺（PPA），以及三种来自新型支架乙二酰胺 - 吡咯酰胺（GPA），吡唑基 - 噻唑（PT）和二苯并 - 硫氮杂-2-酮（DBT）。在生物化学和细胞测定中测试了不同CAM的靶活性和抗病毒功效。分析尺寸排阻色谱和透射电子显微镜显示仅HAP化合物诱导异常非衣壳结构的形成（II类作用模式），而剩余的CAM不影响衣壳大体形态（I类作用模式）。来自HepAD38细胞系的细胞内裂解物，诱导性复制HBV，在用SBA，PPA，GPA，PT或DBT化合物处理后未显示细胞内核心蛋白或衣壳的量减少;然而，HAP治疗导致两者都有显着下降。此外，化合物处理的HepAD38细胞的免疫荧光染色显示，所有非HAP CAM均导致HBV核心抗原（HBcAg）平衡向完全细胞质染色的转变，而HAP诱导HBcAg聚集体在细胞核中的积累。我们的研究表明，新型支架GPA，PT和DBT表现出I类作用模式，类似于SBA和PPA，而HAP仍然是属于II类抑制剂的唯一支架。
关键词：

衣壳;衣壳装配修饰剂;核心蛋白; HBV; Heteroarylpyrimidine

结论：
    30031759
DOI：
    10.1016 / j.antiviral.2018.07.011