Zika virus (ZIKV) infection may be associated with severe complications in fetuses and adults, but treatment options are limited. We performed an in silico structure-based screening of a large chemical library to identify potential ZIKV NS2B-NS3 protease inhibitors. Clinically approved drugs belonging to different drug classes were selected among the 100 primary hit compounds with the highest predicted binding affinities to ZIKV NS2B-NS3-protease for validation studies. ZIKV NS2B-NS3 protease inhibitory activity was validated in most of the selected drugs and in vitro anti-ZIKV activity was identified in two of them (novobiocin and lopinavir-ritonavir). Molecular docking and molecular dynamics simulations predicted that novobiocin bound to ZIKV NS2B-NS3-protease with high stability. Dexamethasone-immunosuppressed mice with disseminated ZIKV infection and novobiocin treatment had significantly (P < 0.05) higher survival rate (100% vs 0%), lower mean blood and tissue viral loads, and less severe histopathological changes than untreated controls. This structure-based drug discovery platform should facilitate the identification of additional enzyme inhibitors of ZIKV.

寨卡病毒（ZIKV）感染可能与胎儿和成人的严重并发症相关，但治疗选择有限。我们对大型化学文库进行了基于计算机模拟的筛选，以识别潜在的ZIKV NS2B-NS3蛋白酶抑制剂。从具有最高预测的对ZIKV NS2B-NS3-蛋白酶的结合亲和力的100种主要命中化合物中选择属于不同药物类别的临床批准药物进行验证研究。ZIKV NS2B-NS3蛋白酶抑制活性已在大多数所选药物中和体外进行了验证在其中的两个中（新霉素和洛匹那韦-利托那韦）鉴定出抗ZIKV活性。分子对接和分子动力学模拟预测，新霉素与ZIKV NS2B-NS3-蛋白酶结合具有很高的稳定性。与未治疗的对照组相比，具有分散的ZIKV感染和新霉素治疗的地塞米松免疫抑制小鼠的存活率显着提高（P  <0.05）（100％vs 0％），平均血液和组织病毒载量较低，组织病理学变化较轻。这个基于结构的药物发现平台应有助于鉴定ZIKV的其他酶抑制剂。