Structure of a dihydroxycoumarin active-site inhibitor in complex with the RNase H domain of HIV-1 reverse transcriptase and structure-activity analysis of inhibitor analogs.
Structure of a dihydroxycoumarin active-site inhibitor in complex with the RNase H domain of HIV-1 reverse transcriptase and structure-activity analysis of inhibitor analogs.
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DOI：
10.1016/j.jmb.2014.05.006
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发表时间：
2014-07-15
影响因子：
5.6
通讯作者：
Arnold E
中科院分区：
文献类型：
--
作者：
Himmel DM;Myshakina NS;Ilina T;Van Ry A;Ho WC;Parniak MA;Arnold E
HIV编码四种基本酶：蛋白酶、整合酶、逆转录酶（RT）相关DNA聚合酶和逆转录酶相关核糖核酸酶H （RNase H）。目前临床批准的抗艾滋病药物靶向除RNase H外的所有HIV酶活性，RNase H已被证明是HIV药物开发的一个非常困难的靶点。我们的高通量筛选活动鉴定了二羟基香豆素化合物F3284-8495是RT RNase H的特异性抑制剂，具有低微摩尔效价。抑制剂与靶标结合的结构信息有助于优化抑制效价。在这里，我们报道了F3284-8495结合到HIV-1 RT分离的RNase H结构域活性位点的晶体结构，分辨率极限为1.71 Å。根据这种结构的预测，得到了具有更好抑制活性的化合物。计算分析表明，结构改变可以提供与RT的额外相互作用，从而提高抑制效力。这些研究证实了F3284-8495可以作为开发HIV RNase H抑制剂的有利化学支架。
HIV encodes four essential enzymes: protease, integrase, reverse transcriptase (RT) associated DNA polymerase, and RT-associated ribonuclease H (RNase H). Current clinically approved anti-AIDS drugs target all HIV enzymatic activities except RNase H, which has proven to be a very difficult target for HIV drug discovery. Our high-throughput screening activities identified the dihydroxycoumarin compound F3284-8495 as a specific inhibitor of RT RNase H, with low micromolar potency in vitro. Optimization of inhibitory potency can be facilitated by structural information about inhibitor-target binding. Here, we report the crystal structure of F3284-8495 bound to the active site of an isolated RNase H domain of HIV-1 RT at a resolution limit of 1.71 Å. From predictions based on this structure, compounds were obtained that showed improved inhibitory activity. Computational analysis suggested structural alterations that could provide additional interactions with RT and thus improve inhibitory potency. These studies established proof-of-concept that F3284-8495 could be used as a favorable chemical scaffold for development of HIV RNase H inhibitors.