Wang Qing-Yin, Dong Hongping, Zou Bin, Karuna Ratna, Wan Kah Fei, Zou Jing, Susila Agatha, Yip Andy, Shan Chao, Yeo Kim Long, Xu Haoying, Ding Mei, Chan Wai Ling, Gu Feng, Seah Peck Gee, Liu Wei, Lakshminarayana Suresh B, Kang CongBao, Lescar Julien, Blasco Francesca, Smith Paul W, Shi Pei-Yong
J Virol. 2015 Aug;89(16):8233-44. doi: 10.1128/JVI.00855-15.
The four serotypes of dengue virus (DENV-1 to -4) represent the most prevalent mosquito-borne viral pathogens in humans. No clinically approved vaccine or antiviral is currently available for DENV. Here we report a spiropyrazolopyridone compound that potently inhibits DENV both in vitro and in vivo. The inhibitor was identified through screening of a 1.8-million-compound library by using a DENV-2 replicon assay. The compound selectively inhibits DENV-2 and -3 (50% effective concentration [EC50], 10 to 80 nM) but not DENV-1 and -4 (EC50,>20 M). Resistance analysis showed that a mutation at amino acid 63 of DENV-2 NS4B (a nonenzymatic transmembrane protein and a component of the viral replication complex) could confer resistance to compound inhibition. Genetic studies demonstrate that variations at amino acid 63 of viral NS4B are responsible for the selective inhibition of DENV-2 and -3. Medicinal chemistry improved the physicochemical properties of the initial “hit” (compound 1), leading to compound 14a, which has good in vivo pharmacokinetics. Treatment of DENV-2-infected AG129 mice with compound 14a suppressed viremia, even when the treatment started after viral infection. The results have proven the concept that inhibitors of NS4B could potentially be developed for clinical treatment of DENV infection. Compound 14a represents a potential preclinical candidate for treatment of DENV-2- and -3-infected patients.
登革病毒的四种血清型(DENV-1至-4)是人类中最常见的蚊媒病毒病原体。目前尚无针对登革病毒的临床批准疫苗或抗病毒药物。在此,我们报告一种螺吡唑并吡啶酮化合物,其在体外和体内均能有效抑制登革病毒。该抑制剂是通过使用DENV-2复制子试验筛选180万种化合物库而鉴定出来的。该化合物选择性抑制DENV-2和-3(50%有效浓度[EC50],10至80 nM),但不抑制DENV-1和-4(EC50,>20 μM)。抗性分析表明,DENV-2 NS4B(一种非酶跨膜蛋白,是病毒复制复合体的一个组成部分)的63位氨基酸突变可赋予对化合物抑制的抗性。遗传学研究表明,病毒NS4B的63位氨基酸变异是DENV-2和-3选择性抑制的原因。药物化学改善了初始“命中”化合物(化合物1)的理化性质,得到了化合物14a,其具有良好的体内药代动力学。用化合物14a治疗DENV-2感染的AG129小鼠可抑制病毒血症,即使在病毒感染后开始治疗也是如此。结果证明了NS4B抑制剂有可能被开发用于登革病毒感染临床治疗的概念。化合物14a是治疗DENV-2和-3感染患者的潜在临床前候选药物。