EASL 2018 SAT-342 由乙肝病毒核心蛋白衍生的钉珠肽 劫持病毒复

StephenW

EASL 2018 SAT-342
The stapled peptides derived from hepatitis B virus core protein
hijack viral replication
J. Han, X. Cong. Peking university people’s hospital, Peking university
hepatology
Email: [email protected]
Background and Aims: Protein-protein interactions (PPI) are
involved in all aspects of the viral lifecycle, including genome
packaging, reverse transcription, intracellular trafficking and cccDNA
epigenetic regulation. Those processes depend on the correct
function and conformation of HBV core protein. To develop PPI
inhibitors targeting core protein is attractive for HBV therapy. Here we
designed a series of stapled peptides based on α-helix domains of
core protein, and completed antiviral activity assay.
Method: Helical peptides with hydrocarbon staples were synthesized,
uptake of peptide by cell and stabilization were assayed.
Antiviral activity was measured by qPCR from HepG2 cells transiently
transfected with four genotype expressing vectors (A∼D). HBsAg and
HBeAg were measured by ELISA in HepDes19 cells. HBV replicative
DNA intermediates and viral RNA were detected by southern and
northern blot analysis. The interaction of stapled peptides with core
protein was determined by co-immunoprecipitation.
Results: Those stapled had more highly levels of cell penetration,
proteolytic resistance, and target affinity than natural α-helix
peptides. Several stapled peptides derived from core protein intradimmer
interface exhibited potent viral load reduction in vitro.
Surprised, Stapled peptides inhibited HBsAg and HBeAg expression
(EC50 50∼100 mM). HBV cccDNA was reduced in HepDeS19 cells
treated with stapled peptides.
Conclusion: Hydrocarbon stapled α-helix peptides derived from core
protein interfere HBV replication, which could be attractive tools for
HBV therapy.

StephenW

EASL 2018 SAT-342
由乙肝病毒核心蛋白衍生的钉珠肽
劫持病毒复制
J. Han，X. Cong。北京大学人民医院，北京大学
肝病
电子邮件：[email protected]
背景和目标：蛋白质 - 蛋白质相互作用（PPI）是
涉及病毒生命周期的各个方面，包括基因组
包装，逆转录，细胞内运输和cccDNA
表观遗传调控。这些进程取决于正确的
HBV核心蛋白的功能和构象。开发PPI
靶向核心蛋白的抑制剂对于HBV治疗有吸引力。在这里，我们
设计了一系列基于α螺旋结构域的钉珠肽
核心蛋白，并完成抗病毒活性测定。
方法：合成具有烃类骨架的螺旋肽，
测定细胞对肽的摄取和稳定性。
通过短暂地从HepG2细胞进行qPCR测量抗病毒活性
用四种基因型表达载体（A〜D）转染。 HBsAg和
在HepDes19细胞中通过ELISA测量HBeAg。 HBV复制
DNA中间体和病毒RNA检测南方和南方
Northern印迹分析。钉珠肽与核心的相互作用
蛋白质通过共免疫沉淀来测定。
结果：钉合的细胞渗透水平更高，
蛋白水解抗性和目标亲和力比天然α-螺旋
肽。来自核心蛋白内切肽的几种钉合肽
界面在体外表现出有效的病毒载量降低。
惊讶的钉装肽抑制了HBsAg和HBeAg的表达
（EC50 50〜100mM）。 HBV cccDNA在HepDeS19细胞中减少
用钉珠肽处理。
结论：来自核心的烃钉合α-螺旋肽
蛋白质干扰HBV复制，这可能是有吸引力的工具
HBV治疗。