High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses.
Identifieur interne : 000537 ( PubMed/Checkpoint ); précédent : 000536; suivant : 000538High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses.
Auteurs : Liang Shen [République populaire de Chine] ; Junwei Niu [République populaire de Chine] ; Chunhua Wang [République populaire de Chine] ; Baoying Huang [République populaire de Chine] ; Wenling Wang [République populaire de Chine] ; Na Zhu [République populaire de Chine] ; Yao Deng [République populaire de Chine] ; Huijuan Wang [République populaire de Chine] ; Fei Ye [République populaire de Chine] ; Shan Cen [République populaire de Chine] ; Wenjie Tan [Oman]Source :
- Journal of virology [ 1098-5514 ] ; 2019.
Abstract
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.
DOI: 10.1128/JVI.00023-19
PubMed: 30918074
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Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Immunology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China tanwj28@163.com.</nlm:affiliation><country wicri:rule="url">Oman</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement 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Shen</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Niu, Junwei" sort="Niu, Junwei" uniqKey="Niu J" first="Junwei" last="Niu">Junwei Niu</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Chunhua" sort="Wang, Chunhua" uniqKey="Wang C" first="Chunhua" last="Wang">Chunhua Wang</name><affiliation wicri:level="3"><nlm:affiliation>National Institutes for Food and Drug Control, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Institutes for Food and Drug Control, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Huang, Baoying" sort="Huang, Baoying" uniqKey="Huang B" first="Baoying" last="Huang">Baoying Huang</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Wenling" sort="Wang, Wenling" uniqKey="Wang W" first="Wenling" last="Wang">Wenling Wang</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Zhu, Na" sort="Zhu, Na" uniqKey="Zhu N" first="Na" last="Zhu">Na Zhu</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Deng, Yao" sort="Deng, Yao" uniqKey="Deng Y" first="Yao" last="Deng">Yao Deng</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Huijuan" sort="Wang, Huijuan" uniqKey="Wang H" first="Huijuan" last="Wang">Huijuan Wang</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Ye, Fei" sort="Ye, Fei" uniqKey="Ye F" first="Fei" last="Ye">Fei Ye</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Cen, Shan" sort="Cen, Shan" uniqKey="Cen S" first="Shan" last="Cen">Shan Cen</name><affiliation wicri:level="3"><nlm:affiliation>Department of Immunology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Immunology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><affiliation wicri:level="3"><nlm:affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China tanwj28@163.com.</nlm:affiliation><country wicri:rule="url">Oman</country><wicri:regionArea>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author></analytic><series><title level="j">Journal of virology</title><idno type="eISSN">1098-5514</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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 <i>Renilla</i> luciferase (rOC43-ns2Del-Rluc) and validated the inhibitors using multiple genetically distinct CoVs <i>in vitro</i> We screened 56 hits from the HTS data and validated 36 compounds <i>in vitro</i> 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 <i>in vitro</i> 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 <i>in vivo</i> 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.<b>IMPORTANCE</b> 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 <i>in vitro</i> 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.</div></front></TEI><pubmed><MedlineCitation Status="In-Data-Review" Owner="NLM"><PMID Version="1">30918074</PMID><DateRevised><Year>2020</Year><Month>02</Month><Day>25</Day></DateRevised><Article PubModel="Electronic-Print"><Journal><ISSN IssnType="Electronic">1098-5514</ISSN><JournalIssue CitedMedium="Internet"><Volume>93</Volume><Issue>12</Issue><PubDate><Year>2019</Year><Month>Jun</Month><Day>15</Day></PubDate></JournalIssue><Title>Journal of virology</Title><ISOAbbreviation>J. Virol.</ISOAbbreviation></Journal><ArticleTitle>High-Throughput Screening and Identification of Potent Broad-Spectrum Inhibitors of Coronaviruses.</ArticleTitle><ELocationID EIdType="pii" ValidYN="Y">e00023-19</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1128/JVI.00023-19</ELocationID><Abstract><AbstractText>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 <i>Renilla</i> luciferase (rOC43-ns2Del-Rluc) and validated the inhibitors using multiple genetically distinct CoVs <i>in vitro</i> We screened 56 hits from the HTS data and validated 36 compounds <i>in vitro</i> 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 <i>in vitro</i> 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 <i>in vivo</i> 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.<b>IMPORTANCE</b> 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 <i>in vitro</i> 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.</AbstractText><CopyrightInformation>Copyright © 2019 American Society for Microbiology.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y" EqualContrib="Y"><LastName>Shen</LastName><ForeName>Liang</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y" EqualContrib="Y"><LastName>Niu</LastName><ForeName>Junwei</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Chunhua</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>National Institutes for Food and Drug Control, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Baoying</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Wenling</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhu</LastName><ForeName>Na</ForeName><Initials>N</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Deng</LastName><ForeName>Yao</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Huijuan</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ye</LastName><ForeName>Fei</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Cen</LastName><ForeName>Shan</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Department of Immunology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tan</LastName><ForeName>Wenjie</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>NHC Key Laboratory of Biosafety, Ministry of Health, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, China tanwj28@163.com.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>05</Month><Day>29</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>J 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