Screening and identification of HTNV<sub>pv</sub> entry inhibitors with high-throughput pseudovirus-based chemiluminescence

Xiaojing Wen; Li Zhang; Qiang Liu; Xinyue Xiao; Weijin Huang; Youchun Wang

doi:10.1016/j.virs.2022.04.015

August 2022

Xiaojing Wen, Li Zhang, Qiang Liu, Xinyue Xiao, Weijin Huang and Youchun Wang. Screening and identification of HTNVpv entry inhibitors with high-throughput pseudovirus-based chemiluminescence[J]. Virologica Sinica, 2022, 37(4): 531-537. doi: 10.1016/j.virs.2022.04.015

Citation: Xiaojing Wen, Li Zhang, Qiang Liu, Xinyue Xiao, Weijin Huang, Youchun Wang. Screening and identification of HTNV_pv entry inhibitors with high-throughput pseudovirus-based chemiluminescence .VIROLOGICA SINICA, 2022, 37(4) : 531-537. http://dx.doi.org/10.1016/j.virs.2022.04.015

基于假病毒的高通量化学发光法筛选和确认汉滩假病毒进入抑制剂

温小菁 ^a,b ,
张黎 ^a ,
刘强 ^a ,
肖新月 ^c ,
黄维金 ^a,, ,
王佑春 ^a,,

1 中国食品药品检定研究院;
2 山东大学

通讯作者： 黄维金, huangweijin@nifdc.org.cn; 王佑春, wangyc@nifdc.org.cn
收稿日期： 2021-03-12
录用日期： 2021-09-03

摘要

汉坦病毒中像汉滩病毒、汉城病毒是引起汉坦病毒心肺综合征和肾综合征出血热的病原体，是重要的人畜共患病原体。中国是肾综合征出血热发病率最高的国家，主要由汉滩病毒和汉城病毒引起。目前还没有批准的抗病毒药物用于治疗这些汉坦病毒疾病。本研究开发了一种基于化学发光的高通量筛选方法筛选汉滩假病毒抑制剂，药物库包含1813份上市药物和556份中药来源小分子化合物。我们筛选到6种化合物在体外具有较高的抗汉滩假病毒活性。（EC50 0.1–2.2μmol/L，SI指数40-900）。筛选到的6中化合物中千金藤素不仅具有较好的体外抗病毒活性，还能抑制汉滩假病毒感染的小鼠，感染后5小时的抑制率分别为94%（180 mg/kg/d，P < 0.01）、93%（90 mg/kg/d，P < 0.01）或92%（45 mg/kg/d，P < 0.01）。添加时间实验表明千金藤素的抗病毒机制主要是膜融合和进入阶段。在本研究中，我们建立了抗病毒药物的高通量筛选方法，并表明千金藤素是治疗汉坦病毒心肺综合征和肾综合征出血热的候选药物。这些发现可能为汉坦病毒感染者的治疗提供一个起点。
- 汉滩病毒
- , 高通量筛选
- , 抑制剂
- , 药物再利用

Screening and identification of HTNV_pv entry inhibitors with high-throughput pseudovirus-based chemiluminescence

Xiaojing Wen ^a,b ,
Li Zhang ^a ,
Qiang Liu ^a ,
Xinyue Xiao ^c ,
Weijin Huang ^a,, ,
Youchun Wang ^a,,

a Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC) and WHO Collaborating Center for Standardization and Evaluation of Biologicals, Beijing 102629, China;
b Department of Microbiological Laboratory Technology, School of Public Health, Shandong University, Jinan 250012, China;

Corresponding author: Weijin Huang, huangweijin@nifdc.org.cn Youchun Wang, wangyc@nifdc.org.cn
Received Date: 12 March 2021
Accepted Date: 03 September 2021

Abstract

Hantaviruses, such as Hantaan virus (HTNV) and Seoul virus, are the causative agents of Hantavirus cardiopulmonary syndrome (HCPS) and hemorrhagic fever with renal syndrome (HFRS), and are important zoonotic pathogens. China has the highest incidence of HFRS, which is mainly caused by HTNV and Seoul virus. No approved antiviral drugs are available for these hantaviral diseases. Here, a chemiluminescence-based high-throughput-screening (HTS) assay was developed and used to screen HTNV pseudovirus (HTNV_pv) inhibitors in a library of 1813 approved drugs and 556 small-molecule compounds from traditional Chinese medicine sources. We identified six compounds with in vitro anti-HTNV_pv activities in the low-micromolar range (EC₅₀ values of 0.1–2.2 μmol/L; selectivity index of 40–900). Among the six selected compounds, cepharanthine not only showed good anti-HTNV_pv activity in vitro but also inhibited HTNV_pv-fluc infection in Balb/c mice 5 h after infection by 94% (180 mg/kg/d, P < 0.01), 93% (90 mg/kg/d, P < 0.01), or 92% (45 mg/kg/d, P < 0.01), respectively, in a bioluminescent imaging mouse model. A time-of-addition analysis suggested that the antiviral mechanism of cepharanthine involves the membrane fusion and entry phases. Overall, we have established a HTS method for antiviral drugs screening, and shown that cepharanthine is a candidate for HCPS and HFRS therapy. These findings may offer a starting point for the treatment of patients infected with hantaviruses.
- Hantaan virus (HTNV)
- , High-throughput screening (HTS)
- , Inhibitor
- , Drug repurposing

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