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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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2019
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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. |
format | Online Article Text |
id | pubmed-6613765 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
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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