Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry
Antiviral targeting of virus envelope proteins is an effective strategy for therapeutic intervention of viral infections. Here, we took a computer-guided approach with the aim of identifying new antivirals against the envelope protein E2 of bovine viral diarrhea virus (BVDV). BVDV is an enveloped vi...
| Main Authors: | , , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
Elsevier
2019
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/4480 https://www.sciencedirect.com/science/article/pii/S0166354217305983?via%3Dihub#! https://doi.org/10.1016/j.antiviral.2017.10.010 |
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| author | Pascual, María José Merwaiss, Fernando Leal, Emilse Soledad Quintana, Maria Eugenia Capozzo, Alejandra Victoria Cavasotto, Claudio Norberto Bollini, Mariela Alvarez, Diego Ezequiel |
| author_browse | Alvarez, Diego Ezequiel Bollini, Mariela Capozzo, Alejandra Victoria Cavasotto, Claudio Norberto Leal, Emilse Soledad Merwaiss, Fernando Pascual, María José Quintana, Maria Eugenia |
| author_facet | Pascual, María José Merwaiss, Fernando Leal, Emilse Soledad Quintana, Maria Eugenia Capozzo, Alejandra Victoria Cavasotto, Claudio Norberto Bollini, Mariela Alvarez, Diego Ezequiel |
| author_sort | Pascual, María José |
| collection | INTA Digital |
| description | Antiviral targeting of virus envelope proteins is an effective strategy for therapeutic intervention of viral infections. Here, we took a computer-guided approach with the aim of identifying new antivirals against the envelope protein E2 of bovine viral diarrhea virus (BVDV). BVDV is an enveloped virus with an RNA genome responsible for major economic losses of the cattle industry worldwide. Based on the crystal structure of the envelope protein E2, we defined a binding site at the interface of the two most distal domains from the virus membrane and pursued a hierarchical docking-based virtual screening search to identify small-molecule ligands of E2. Phenyl thiophene carboxamide derivative 12 (PTC12) emerged as a specific inhibitor of BVDV replication from in vitro antiviral activity screening of candidate molecules, displaying an IC50 of 0.30 μM against the reference NADL strain of the virus. Using reverse genetics we constructed a recombinant BVDV expressing GFP that served as a sensitive reporter for the study of the mechanism of action of antiviral compounds. Time of drug addition assays showed that PTC12 inhibited an early step of infection. The mechanism of action was further dissected to find that the compound specifically acted at the internalization step of virus entry. Interestingly, we demonstrated that similar to PTC12, the benzimidazole derivative 03 (BI03) selected in the virtual screen also inhibited internalization of BVDV. Furthermore, docking analysis of PTC12 and BI03 into the binding site revealed common interactions with amino acid residues in E2 suggesting that both compounds could share the same molecular target. In conclusion, starting from a targeted design strategy of antivirals against E2 we identified PTC12 as a potent inhibitor of BVDV entry. The compound can be valuable in the design of antiviral strategies in combination with already well-characterized polymerase inhibitors of BVDV. |
| format | Artículo |
| id | INTA4480 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA44802019-02-21T14:02:16Z Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry Pascual, María José Merwaiss, Fernando Leal, Emilse Soledad Quintana, Maria Eugenia Capozzo, Alejandra Victoria Cavasotto, Claudio Norberto Bollini, Mariela Alvarez, Diego Ezequiel Pestivirus de la Diarrea Bovina Inhibición Viricidas Bovine Diarrhoea Pestivirus Inhibition Antiviral Agents BVDV Protein E2 Proteína E2 Antiviral targeting of virus envelope proteins is an effective strategy for therapeutic intervention of viral infections. Here, we took a computer-guided approach with the aim of identifying new antivirals against the envelope protein E2 of bovine viral diarrhea virus (BVDV). BVDV is an enveloped virus with an RNA genome responsible for major economic losses of the cattle industry worldwide. Based on the crystal structure of the envelope protein E2, we defined a binding site at the interface of the two most distal domains from the virus membrane and pursued a hierarchical docking-based virtual screening search to identify small-molecule ligands of E2. Phenyl thiophene carboxamide derivative 12 (PTC12) emerged as a specific inhibitor of BVDV replication from in vitro antiviral activity screening of candidate molecules, displaying an IC50 of 0.30 μM against the reference NADL strain of the virus. Using reverse genetics we constructed a recombinant BVDV expressing GFP that served as a sensitive reporter for the study of the mechanism of action of antiviral compounds. Time of drug addition assays showed that PTC12 inhibited an early step of infection. The mechanism of action was further dissected to find that the compound specifically acted at the internalization step of virus entry. Interestingly, we demonstrated that similar to PTC12, the benzimidazole derivative 03 (BI03) selected in the virtual screen also inhibited internalization of BVDV. Furthermore, docking analysis of PTC12 and BI03 into the binding site revealed common interactions with amino acid residues in E2 suggesting that both compounds could share the same molecular target. In conclusion, starting from a targeted design strategy of antivirals against E2 we identified PTC12 as a potent inhibitor of BVDV entry. The compound can be valuable in the design of antiviral strategies in combination with already well-characterized polymerase inhibitors of BVDV. Instituto de Virología Fil: Pascual, María José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Merwaiss, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina Fil: Leal, Emilse Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias ; Argentina Fil: Quintana, Maria Eugenia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Capozzo, Alejandra Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina Fil: Cavasotto, Claudio Norberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Bollini, Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias ; Argentina Dil: Alvarez, Diego Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina 2019-02-21T13:54:01Z 2019-02-21T13:54:01Z 2018-01 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/4480 https://www.sciencedirect.com/science/article/pii/S0166354217305983?via%3Dihub#! 0166-3542 https://doi.org/10.1016/j.antiviral.2017.10.010 eng info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Antiviral research 149 : 179-190. (January 2018) |
| spellingShingle | Pestivirus de la Diarrea Bovina Inhibición Viricidas Bovine Diarrhoea Pestivirus Inhibition Antiviral Agents BVDV Protein E2 Proteína E2 Pascual, María José Merwaiss, Fernando Leal, Emilse Soledad Quintana, Maria Eugenia Capozzo, Alejandra Victoria Cavasotto, Claudio Norberto Bollini, Mariela Alvarez, Diego Ezequiel Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| title | Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| title_full | Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| title_fullStr | Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| title_full_unstemmed | Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| title_short | Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| title_sort | structure based drug design for envelope protein e2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry |
| topic | Pestivirus de la Diarrea Bovina Inhibición Viricidas Bovine Diarrhoea Pestivirus Inhibition Antiviral Agents BVDV Protein E2 Proteína E2 |
| url | http://hdl.handle.net/20.500.12123/4480 https://www.sciencedirect.com/science/article/pii/S0166354217305983?via%3Dihub#! https://doi.org/10.1016/j.antiviral.2017.10.010 |
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