Novel Benzoxazole Inhibitor of Dengue Virus Replication That Targets the NS3 Helicase
Novel Benzoxazole Inhibitor of Dengue Virus Replication That Targets the NS3 Helicase
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DOI：
10.1128/aac.02251-12
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发表时间：
2013-04-01
影响因子：
4.9
通讯作者：
Jordan, Robert
中科院分区：
文献类型：
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作者：
Byrd, Chelsea M.;Grosenbach, Douglas W.;Jordan, Robert
登革热病毒（DENV）是主要的蚊媒病毒性病原体，每年全世界估计有5000万至1亿人感染。在过去50年中，登革热的发病率急剧增加，该病毒目前在100多个国家流行。此外，现在在同一地理区域发现了DENV的多种血清型，增加了出现更严重疾病形式的可能性。尽管进行了广泛的研究，但仍然没有批准的疫苗或商业上可用的治疗DENV感染的疗法。在这里，我们报告了使用全病毒试验对化合物文库进行高通量筛选的结果，该结果鉴定出一种新的DENV小分子抑制剂ST-610，该抑制剂在体外有效且选择性地抑制所有四种DENV血清型的复制。对耐药病毒分离株的序列分析发现，在NS3解旋酶结构域存在一个单点突变A263T，使其对该化合物产生耐药性。ST-610在分子信标解旋酶实验中抑制DENV NS3解旋酶RNA解绕活性，但在孔雀石绿atp酶实验中不抑制核苷三磷酸酶活性。ST-610是非诱变的，在小鼠中具有良好的耐受性（无毒），并且在DENV感染的亚致死小鼠模型中显示出有效性，与对照物相比，能够显著降低病毒血症和病毒载量。
Dengue virus (DENV) is the predominant mosquito-borne viral pathogen that infects humans with an estimated 50 to 100 million infections per year worldwide. Over the past 50 years, the incidence of dengue disease has increased dramatically and the virus is now endemic in more than 100 countries. Moreover, multiple serotypes of DENV are now found in the same geographic region, increasing the likelihood of more severe forms of disease. Despite extensive research, there are still no approved vaccines or therapeutics commercially available to treat DENV infection. Here we report the results of a high-throughput screen of a chemical compound library using a whole-virus assay that identified a novel small-molecule inhibitor of DENV, ST-610, that potently and selectively inhibits all four serotypes of DENV replication in vitro. Sequence analysis of drug-resistant virus isolates has identified a single point mutation, A263T, in the NS3 helicase domain that confers resistance to this compound. ST-610 inhibits DENV NS3 helicase RNA unwinding activity in a molecular-beacon-based helicase assay but does not inhibit nucleoside triphosphatase activity based on a malachite green ATPase assay. ST-610 is nonmutagenic, is well tolerated (nontoxic) in mice, and has shown efficacy in a sublethal murine model of DENV infection with the ability to significantly reduce viremia and viral load compared to vehicle controls.