APASL2017 [OP112] 结合HBx的化合物抑制HBV DNA 复制

StephenW

APASL2017[OP112]
Chemical compounds that bind to HBx inhibit HBV DNA
replication
Sayuri Morimoto1, Ryosuke Muroyama1, Yasuo Tanaka2, Sayaka
Ito1, Ryo Nakagawa1, Kaku Goto1, Jun Arai1, Yoshimi Kaise1
1The Institute of Medical Science, The University of Tokyo, Tokyo,
Japan; 2Department of Gastroenterology, Graduate School of
Medicine, The University of Tokyo, Tokyo, Japan
Background: Multifunctional protein HBx is associated with HBV
replication and carcinogenesis. Chemical compounds that suppress
the function of HBx by binding to it could elucidate the mechanism of
HBx-dependent HBV replication and carcinogenesis. Their modes of
action being completely different from that of nucleoside analogues
or IFN treatment, they would provide a novel means to treat HBVrelated
diseases.
Methods: GST-tagged HBx was expressed and purified in E. coli.
The library of 1018 FDA-approved drugs was screened for the
binding to HBx by surface plasmon resonance analysis. The compounds’
effects on carcinogenesis were measured by cell proliferation
assay using Hep3B cells that demonstrate HBx-dependent proliferation
and Huh7 cells that do not contain integrated HBV DNA. The
compounds’ effects on HBV replication were measured by the
amount of HBV DNA in the culture medium using real-time PCR. In
this experiment, HepG2.2.15.7 cells having stable HBV expression
were used.
Result: Of the 1018 FDA-approved drugs in the library, 22 were
found to bind to HBx, and 6 of them could strongly bind to HBx
compared to others. These six compounds were further tested and cell
proliferation assay revealed that there was no difference in the rate of
cell proliferation between Hep3B and Huh7 cell lines. On the other
hand, two compounds showed more than 50% inhibition of the
amount of HBV DNA. One of them showed more than 90% inhibition,
almost as good as 95% inhibition of nucleoside analogue
entecavir.
Conclusion: Some FDA-approved drugs that bind to HBx were found
to inhibit HBV DNA replication. These compounds, working in a
different manner than entecavir, could be an attractive option for the
treatment of HBV-related diseases.

StephenW

APASL2017 [OP112]
结合HBx的化合物抑制HBV DNA
复制
Sayuri Morimoto1，Ryosuke Muroyama1，Yasuo Tanaka2，Sayaka
Ito1，Ryo Nakagawa1，Kaku Goto1，Jun Arai1，Yoshimi Kaise1
1东京大学医学科学研究所，东京，
日本;西北大学大学院医学研究科
医学，东京大学，东京，日本
背景：多功能蛋白HBx与HBV相关
复制和癌发生。抑制的化合物
通过结合HBx的功能可以阐明其机制
HBx依赖性HBV复制和癌发生。他们的模式
作用与核苷类似物完全不同
或IFN治疗，它们将提供治疗HBV相关的新方法
疾病。
方法：在大肠杆菌中表达和纯化GST-标记的HBx。
筛选了1018个FDA批准的药物库
通过表面等离振子共振分析结合HBx。化合物
通过细胞增殖测量对癌发生的作用
测定使用展示HBx依赖增殖的Hep3B细胞
和不含有整合的HBV DNA的Huh7细胞。的
化合物对HBV复制的影响
使用实时PCR在培养基中的HBV DNA的量。在
本实验中，HepG2.2.15.7细胞具有稳定的HBV表达
。
结果：在图书馆的1018个FDA批准的药物中，22个
发现与HBx结合，其中6个可以与HBx强烈结合
相比其他。这六种化合物进一步测试和细胞
增殖测定显示其比率无差异
Hep3B和Huh7细胞之间的细胞增殖。在另一
手，两种化合物显示超过50％的抑制
HBV DNA量。其中一个显示超过90％的抑制，
几乎达到95％的核苷类似物抑制
恩替卡韦。
结论：发现了一些与HBx结合的FDA批准的药物
以抑制HBV DNA复制。这些化合物，在
不同于恩替卡韦，可能是一个有吸引力的选择
治疗HBV相关疾病。

MP4

不知是哪个药