Inhibition of hepatitis C viral RNA-dependent RNA polymerase by α-P-boranophosphate nucleotides: exploring a potential strategy for mechanism-based HCV drug design.
Inhibition of hepatitis C viral RNA-dependent RNA polymerase by α-P-boranophosphate nucleotides: exploring a potential strategy for mechanism-based HCV drug design.
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通过α-P-硼磷酸核苷酸抑制丙型肝炎病毒RNA依赖性RNA聚合酶：探索基于机制的HCV药物设计的潜在策略。
DOI：
10.1016/j.antiviral.2013.02.014
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发表时间：
2013-05
影响因子：
7.6
通讯作者：
Shaw, Barbara Ramsay
中科院分区：
文献类型：
--
作者：
Cheek, Marcus Adrian;Sharaf, Mariam L.;Dobrikov, Mikhail I.;Shaw, Barbara Ramsay
关键词：
改善慢性 HCV 感染的治疗方法仍然是一个挑战，需要新的化学策略来扩展当前的范例。 HCV RNA 聚合酶 (RdRP) 一直是抗病毒药物开发的目标。我们首次表明，硼烷磷酸盐 (BP) 修饰提高了 ATP 类似物进入 HCV NS5BΔ55 RdRP 催化 RNA 的底物效率。硼磷酸核苷酸含有硼烷 (BH3) 基团，该基团取代了正常磷酸基团的非桥联磷酰氧，从而形成一类能够调节遗传信息读写的修饰等电子 DNA 和 RNA 模拟物。我们确定 HCV NS5BΔ55 是一种立体特异性酶，包含 ATPαB 和两种硼磷酸类似物的 Rp 异构体：2'-O-甲基腺苷 5'-(α-P-borano) 三磷酸 (2'-OMe ATPαB, 5a) 和 3'-脱氧腺苷 5'-(α-P-borano) 三磷酸（3'-dATPαB，5b）。 ATPαB (6) 的 Rp 非对映异构体没有核糖修饰，被发现是比天然 ATP 稍好的底物，表观米氏常数 (Km) 降低了 42%。硼磷酸盐修饰后，2'-O-Me 和 3'-脱氧 ATP 的 IC50 均降低了 16 倍。通过测定稳态抑制常数 (Ki) 进一步证实了这种“硼拉诺效应”，显示出可比较的效力变化（21 倍）。这些实验还表明硼磷酸盐类似物5a和5b通过竞争性抑制模式抑制HCV NS5B。这一证据与之前的晶体结构数据一起，进一步支持了这样的观点：HCV NS5B（以与 HIV-1 RT 类似的方式）通过失去核糖上的 3’-OH 和活性位点残基之间的相互作用，或在磷酰基转移过程中失去 3’-OH 和焦磷酸离去基团之间的分子内氢键相互作用来区分 3’-脱氧修饰。据我们所知，这些数据代表了首次研究磷酸盐修饰的 NTP 作为 HCV NS5B RdRP 的底物。
Improved treatments for chronic HCV infections remain a challenge, and new chemical strategies are needed to expand the current paradigm. The HCV RNA polymerase (RdRP) has been a target for antiviral development. For the first time we show that the boranophosphate (BP) modification increases the substrate efficiency of ATP analogs into HCV NS5BΔ55 RdRP-catalyzed RNA. Boranophosphate nucleotides contain a borane (BH3) group substituted for a non-bridging phosphoryl oxygen of a normal phosphate group, resulting in a class of modified isoelectronic DNA and RNA mimics capable of modulating the reading and writing of genetic information. We determine that HCV NS5BΔ55, being a stereospecific enzyme, incorporates the Rp isomer of both ATPαB and the two boranophosphate analogs: 2’-O-methyladenosine 5’-(α-P-borano) triphosphate (2’-OMe ATPαB, 5a) and 3’-deoxyadenosine 5’-(α-P-borano) triphosphate (3’-dATPαB, 5b). The Rp diastereomer of ATPαB (6), having no ribose modifications, was found to be a slightly better substrate than natural ATP, showing a 42% decrease in the apparent Michaelis-Menten constant (Km). The IC50 of both 2’-O-Me and 3’-deoxy ATP was decreased with the boranophosphate modification up to 16-fold. This “borano effect” was further confirmed by determining the steady-state inhibitory constant (Ki), showing a comparable potency shift (21-fold). These experiments also indicate that the boranophosphate analogs 5a and 5b inhibit HCV NS5B through a competitive mode of inhibition. This evidence, together with previous crystal structure data, further supports the idea that HCV NS5B (in a similar manner to HIV-1 RT) discriminates against the 3’-deoxy modification via lost interactions between the 3’-OH on the ribose and the active site residues, or lost intramolecular hydrogen bonding interactions between the 3’-OH and the pyrophosphate leaving group during phosphoryl transfer. To our knowledge, these data represent the first time a phosphate modified NTP has been studied as a substrate for HCV NS5B RdRP.
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影响因子：
13.5
作者：
Ghany, Marc G.;Nelson, David R.;Strader, Doris B.;Thomas, David L.;Seeff, Leonard B.
通讯作者：
Seeff, Leonard B.
影响因子：
5.4
作者：
Bressanelli, S;Tomei, L;De Francesco, R
通讯作者：
De Francesco, R
影响因子：
11.4
作者：
Meyer, P;Schneider, B;Canard, B
通讯作者：
Canard, B
DOI：
10.1074/jbc.m111.325530
发表时间：
2012-03-23
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Jin Z;Leveque V;Ma H;Johnson KA;Klumpp K
通讯作者：
Klumpp K
影响因子：
4.9
作者：
Carroll, Steven S.;Ludmerer, Steven;Olsen, David B.
通讯作者：
Olsen, David B.