Zhou Zhigang, Khaliq Mansoora, Suk Jae-Eun, Patkar Chinmay, Li Long, Kuhn Richard J, Post Carol Beth
Department of Medicinal Chemistry and Molecular Pharmacology, Markey Center for Structural Biology and Purdue Cancer Center, Purdue University, West Lafayette, Indiana 47907, USA.
ACS Chem Biol. 2008 Dec 19;3(12):765-75. doi: 10.1021/cb800176t.
Infection by the mosquito-borne dengue virus causes dengue fever and the sometimes fatal dengue hemorrhagic fever. The increasing number of dengue infections per year suggests that the virus is becoming more virulent and its transmission is expanding. Nevertheless, no effective treatment for dengue infection currently exists. In a search for antiviral agents effective against dengue virus, we investigated the potential of targeting a structural protein site rather than an enzymatic one. Using this approach, we now report the discovery of a small molecule ligand that inhibits viral growth. Our results also provide the first evidence that the binding site, a pocket located at the hinge between domains 1 and 2 of the envelope protein (E protein) on the virus surface, is a valid target for antiviral therapy. Ligand candidates were identified from libraries of approximately 142,000 compounds using a computational high-throughput screening protocol targeting this pocket of the E protein. Cell-based assays were conducted on 23 top-ranked compounds. Among four with good antiviral activity profiles, the compound P02 was found to inhibit viral reproduction at micromolar concentrations. Using saturation transfer difference NMR spectroscopy, we also show that the compound binds virus and competes for binding E protein with the known ligand N-octyl-beta-D-glucoside. Together, the results are consistent with an inhibition mechanism against maturation or host-cell entry mediated by ligand binding to the E-protein pocket. P02 is a promising lead compound for future development of an effective treatment against dengue virus and related flaviviruses.
由蚊子传播的登革病毒感染会引发登革热以及有时会致命的登革出血热。每年登革感染病例数的增加表明该病毒的毒性正在增强，其传播范围也在扩大。然而，目前尚无针对登革感染的有效治疗方法。在寻找有效对抗登革病毒的抗病毒药物的过程中，我们研究了靶向一种结构蛋白位点而非酶类位点的潜力。采用这种方法，我们现在报告发现了一种抑制病毒生长的小分子配体。我们的结果还首次证明，位于病毒表面包膜蛋白（E蛋白）结构域1和结构域2之间铰链处的一个口袋状结合位点，是抗病毒治疗的一个有效靶点。使用针对E蛋白这个口袋的计算高通量筛选方案，从大约142,000种化合物的文库中鉴定出了候选配体。对23种排名靠前的化合物进行了基于细胞的检测。在四种具有良好抗病毒活性的化合物中，发现化合物P02在微摩尔浓度下就能抑制病毒繁殖。利用饱和转移差核磁共振光谱技术，我们还表明该化合物能与病毒结合，并与已知配体N-辛基-β-D-葡萄糖苷竞争结合E蛋白。这些结果共同表明，其抑制机制是通过配体与E蛋白口袋结合介导的对病毒成熟或进入宿主细胞的抑制。P02是未来开发有效对抗登革病毒及相关黄病毒治疗方法的一种有前景的先导化合物。
