EASL 2018 SAT-355 乙型肝炎病毒肽阵列展示候选机制 CRV431抗HBV活

StephenW

EASL 2018 SAT-355
HBV peptide array demonstrates candidate mechanisms of
CRV431 anti-HBV activity
R. Foster1, A. Levin2, C. Le2, D. Ure1, D. Trepanier1, L. Tyrrell2. 1ContraVir
Pharmaceuticals, Inc., Drug Development, Edison, United States; 2Li Ka
Shing Institute of Virology, Department of Medical Microbiology and
Immunology, Edmonton, Canada
Email: [email protected]
Background and Aims: CRV431 is a cyclosporine A analog that
reduces HBsAg, HBV DNA, and other HBV markers. Most of the
CRV431 activities are thought to occur by blocking participation of
cellular cyclophilins, the primary molecular target of CRV431, in the
HBV life cycle. However, the specific cyclophilin interactions that
influence HBV have not yet been identified. The aims of the current
study were to determine whether cyclophilin A can bind directly to
HBV proteins, whether CRV431 blocks the interactions, and to
identify the specific HBV protein sequences to which cyclophilin
binds.
Method: Soluble, recombinant cyclophilin A was applied to immobilized,
15-amino acid, overlapping peptides representing the entire
HBV proteome, and cyclophilin A binding to the peptides was
detected with cyclophilin A antibody. CRV431 was additionally
applied in some experiments to block the binding. The sequences
of the peptides that bound to cyclophilin A were analyzed in detail,
and soluble peptides were used in competition experiments to
validate cyclophilin A binding to the HBV proteins.
Results: Cyclophilin A bound to 10 HBV-derived peptides, and all 10
binding events were inhibited by CRV431. The peptides that bound
cyclophilin A were derived from preS1 HBsAg, HBV polymerase,
precore, and core antigen. Furthermore, the peptides overlapped with
sequences implicated in regulation of polymerase nuclear import,
HBsAg transport and secretion, and capsid formation. One additional
interaction between cyclophilin A and a polymerase-derived peptide
was found but only in the presence of CRV431. Current studies are
investigating the physiological relevance of the cyclophilin A binding
events observed in the peptide arrays.
Conclusion: These studies identified multiple cyclophilin-HBV
interactions that may be the target(s) of CRV431 action.

StephenW

EASL 2018 SAT-355
乙型肝炎病毒肽阵列展示候选机制
CRV431抗HBV活性
R.Foster1，A.Levin2，C.Le2，D.Ure1，D.Turpanier1，L.Tyrrell2。 1ContraVir
Pharmaceuticals，Inc.，Drug Development，Edison，United States; 2李嘉
盛诚医学微生物学与病毒学研究所
免疫学，加拿大埃德蒙顿
电子邮件：[email protected]
背景和目标：CRV431是一种环孢素A类似物
减少HBsAg，HBV DNA和其他HBV标记。大部分的
CRV431活动被认为是通过阻止参与而发生的
细胞亲环素，CRV431的主要分子靶点
HBV生命周期。但是，具体的亲环蛋白相互作用
影响HBV尚未确定。目前的目标
研究确定亲环蛋白A是否可以直接结合到
乙肝病毒蛋白，无论是CRV431阻止相互作用，并
鉴定亲环蛋白的特定HBV蛋白序列
结合。
方法：将可溶性重组亲环素A用于固定化，
15个氨基酸，代表整个的重叠肽
HBV蛋白质组和亲环蛋白A与肽的结合
用亲环素A抗体检测。另外还有CRV431
在一些实验中应用来阻断结合。序列
结合亲环蛋白A的肽进行了详细的分析，
并将可溶性肽用于竞争实验中
验证亲环蛋白A与HBV蛋白的结合。
结果：亲环蛋白A结合10个HBV衍生肽，全部10个
结合事件被CRV431抑制。结合的肽
亲环蛋白A来源于preS1 HBsAg，HBV聚合酶，
前核心和核心抗原。此外，肽重叠
涉及聚合酶核进口调控的序列，
HBsAg转运和分泌以及衣壳形成。另外一个
亲环蛋白A和聚合酶衍生肽之间的相互作用
被发现，但只有在CRV431的存在。目前的研究是
研究亲环蛋白A结合的生理相关性
在肽阵列中观察到的事件。
结论：这些研究确定了多种亲环蛋白-HBV
相互作用可能是CRV431行动的目标。