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High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses

Coronaviruses (CoVs) act as cross-species viruses and have the potential to spread rapidly into new host species and cause epidemic diseases. Despite the severe public health threat of severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome CoV (MERS-CoV), there are curren...

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Autores principales: Shen, Liang, Niu, Junwei, Wang, Chunhua, Huang, Baoying, Wang, Wenling, Zhu, Na, Deng, Yao, Wang, Huijuan, Ye, Fei, Cen, Shan, Tan, Wenjie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6613765/
https://www.ncbi.nlm.nih.gov/pubmed/30918074
http://dx.doi.org/10.1128/JVI.00023-19
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author Shen, Liang
Niu, Junwei
Wang, Chunhua
Huang, Baoying
Wang, Wenling
Zhu, Na
Deng, Yao
Wang, Huijuan
Ye, Fei
Cen, Shan
Tan, Wenjie
author_facet Shen, Liang
Niu, Junwei
Wang, Chunhua
Huang, Baoying
Wang, Wenling
Zhu, Na
Deng, Yao
Wang, Huijuan
Ye, Fei
Cen, Shan
Tan, Wenjie
author_sort Shen, Liang
collection PubMed
description Coronaviruses (CoVs) act as cross-species viruses and have the potential to spread rapidly into new host species and cause epidemic diseases. Despite the severe public health threat of severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome CoV (MERS-CoV), there are currently no drugs available for their treatment; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are urgently needed. To search for effective inhibitory agents, we performed high-throughput screening (HTS) of a 2,000-compound library of approved drugs and pharmacologically active compounds using the established genetically engineered human CoV OC43 (HCoV-OC43) strain expressing Renilla luciferase (rOC43-ns2Del-Rluc) and validated the inhibitors using multiple genetically distinct CoVs in vitro. We screened 56 hits from the HTS data and validated 36 compounds in vitro using wild-type HCoV-OC43. Furthermore, we identified seven compounds (lycorine, emetine, monensin sodium, mycophenolate mofetil, mycophenolic acid, phenazopyridine, and pyrvinium pamoate) as broad-spectrum inhibitors according to their strong inhibition of replication by four CoVs in vitro at low-micromolar concentrations. Additionally, we found that emetine blocked MERS-CoV entry according to pseudovirus entry assays and that lycorine protected BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This represents the first demonstration of in vivo real-time bioluminescence imaging to monitor the effect of lycorine on the spread and distribution of HCoV-OC43 in a mouse model. These results offer critical information supporting the development of an effective therapeutic strategy against CoV infection. IMPORTANCE Currently, there is no approved therapy to treat coronavirus infection; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are needed. Based on our high-throughput screening assay using a compound library, we identified seven compounds with broad-spectrum efficacy against the replication of four CoVs in vitro. Additionally, one compound (lycorine) was found to protect BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This inhibitor might offer promising therapeutic possibilities for combatting novel CoV infections in the future.
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spelling pubmed-66137652019-11-29 High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses Shen, Liang Niu, Junwei Wang, Chunhua Huang, Baoying Wang, Wenling Zhu, Na Deng, Yao Wang, Huijuan Ye, Fei Cen, Shan Tan, Wenjie J Virol Vaccines and Antiviral Agents Coronaviruses (CoVs) act as cross-species viruses and have the potential to spread rapidly into new host species and cause epidemic diseases. Despite the severe public health threat of severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome CoV (MERS-CoV), there are currently no drugs available for their treatment; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are urgently needed. To search for effective inhibitory agents, we performed high-throughput screening (HTS) of a 2,000-compound library of approved drugs and pharmacologically active compounds using the established genetically engineered human CoV OC43 (HCoV-OC43) strain expressing Renilla luciferase (rOC43-ns2Del-Rluc) and validated the inhibitors using multiple genetically distinct CoVs in vitro. We screened 56 hits from the HTS data and validated 36 compounds in vitro using wild-type HCoV-OC43. Furthermore, we identified seven compounds (lycorine, emetine, monensin sodium, mycophenolate mofetil, mycophenolic acid, phenazopyridine, and pyrvinium pamoate) as broad-spectrum inhibitors according to their strong inhibition of replication by four CoVs in vitro at low-micromolar concentrations. Additionally, we found that emetine blocked MERS-CoV entry according to pseudovirus entry assays and that lycorine protected BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This represents the first demonstration of in vivo real-time bioluminescence imaging to monitor the effect of lycorine on the spread and distribution of HCoV-OC43 in a mouse model. These results offer critical information supporting the development of an effective therapeutic strategy against CoV infection. IMPORTANCE Currently, there is no approved therapy to treat coronavirus infection; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are needed. Based on our high-throughput screening assay using a compound library, we identified seven compounds with broad-spectrum efficacy against the replication of four CoVs in vitro. Additionally, one compound (lycorine) was found to protect BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This inhibitor might offer promising therapeutic possibilities for combatting novel CoV infections in the future. American Society for Microbiology 2019-05-29 /pmc/articles/PMC6613765/ /pubmed/30918074 http://dx.doi.org/10.1128/JVI.00023-19 Text en Copyright © 2019 American Society for Microbiology. This article is made available via the PMC Open Access Subset for unrestricted noncommercial re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Vaccines and Antiviral Agents
Shen, Liang
Niu, Junwei
Wang, Chunhua
Huang, Baoying
Wang, Wenling
Zhu, Na
Deng, Yao
Wang, Huijuan
Ye, Fei
Cen, Shan
Tan, Wenjie
High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses
title High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses
title_full High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses
title_fullStr High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses
title_full_unstemmed High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses
title_short High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses
title_sort high-throughput screening and identification of potent broad-spectrum inhibitors of coronaviruses
topic Vaccines and Antiviral Agents
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6613765/
https://www.ncbi.nlm.nih.gov/pubmed/30918074
http://dx.doi.org/10.1128/JVI.00023-19
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