Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro
This article evaluated the in vitro antiviral effect of atorvastatin (ATV) against SARS-CoV-2 and identified the interaction affinity between this compound and two SARS-CoV-2 proteins. The antiviral activity of atorvastatin against this virus was evaluated by three different treatment strategies [(i...
- Autores:
-
Zapata-Cardona, María I
Flórez-Álvarez, Lizdany
Hernández López, Juan Carlos
Zapata Builes, Wildeman
Guerra-Sandoval, Ariadna L
Hincapié-García, Jaime
Rugeles, Maria T
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2022
- Institución:
- Universidad Cooperativa de Colombia
- Repositorio:
- Repositorio UCC
- Idioma:
- OAI Identifier:
- oai:repository.ucc.edu.co:20.500.12494/45190
- Acceso en línea:
- https://hdl.handle.net/20.500.12494/45190
- Palabra clave:
- SARS-CoV-2
antiviral
molecular docking
COVID-19
variants
- Rights
- openAccess
- License
- Atribución
| id |
COOPER2_d0a6dd27f06d8ba4f7fbf3d18176da48 |
|---|---|
| oai_identifier_str |
oai:repository.ucc.edu.co:20.500.12494/45190 |
| network_acronym_str |
COOPER2 |
| network_name_str |
Repositorio UCC |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| title |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| spellingShingle |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro SARS-CoV-2 antiviral molecular docking COVID-19 variants |
| title_short |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| title_full |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| title_fullStr |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| title_full_unstemmed |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| title_sort |
Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro |
| dc.creator.fl_str_mv |
Zapata-Cardona, María I Flórez-Álvarez, Lizdany Hernández López, Juan Carlos Zapata Builes, Wildeman Guerra-Sandoval, Ariadna L Hincapié-García, Jaime Rugeles, Maria T |
| dc.contributor.author.none.fl_str_mv |
Zapata-Cardona, María I Flórez-Álvarez, Lizdany Hernández López, Juan Carlos Zapata Builes, Wildeman Guerra-Sandoval, Ariadna L Hincapié-García, Jaime Rugeles, Maria T |
| dc.subject.spa.fl_str_mv |
SARS-CoV-2 antiviral molecular docking COVID-19 variants |
| topic |
SARS-CoV-2 antiviral molecular docking COVID-19 variants |
| description |
This article evaluated the in vitro antiviral effect of atorvastatin (ATV) against SARS-CoV-2 and identified the interaction affinity between this compound and two SARS-CoV-2 proteins. The antiviral activity of atorvastatin against this virus was evaluated by three different treatment strategies [(i) pre-post treatment, (ii) pre-infection treatment, and (iii) post-infection treatment] using Vero E6 and Caco-2 cells. The interaction of atorvastatin with RdRp (RNA-dependent RNA polymerase) and 3CL protease (3-chymotrypsin-like protease) was evaluated by molecular docking. The CC50s (half-maximal cytotoxic concentrations) obtained for ATV were 50.3 and 64.5 μM in Vero E6 and Caco-2, respectively. This compound showed antiviral activity against SARS-CoV-2 D614G strain in Vero E6 with median effective concentrations (EC50s) of 15.4, 12.1, and 11.1μM by pre-post, pre-infection, and post-infection treatments, respectively. ATV also inhibited Delta and Mu variants by pre-post treatment (EC50s of 16.8 and 21.1μM, respectively). In addition, ATV showed an antiviral effect against the D614G strain independent of the cell line (EC50 of 7.4μM in Caco-2). The interaction of atorvastatin with SARS-CoV-2 RdRp and 3CL protease yielded a binding affinity of −6.7kcal/mol and −7.5kcal/mol, respectively. Our study demonstrated the in vitro antiviral activity of atorvastatin against the ancestral SARS-CoV-2 D614G strain and two emerging variants (Delta and Mu), with an independent effect of the cell line. A favorable binding affinity between ATV and viral proteins by bioinformatics methods was found. Due to the extensive clinical experience of atorvastatin use, it could prove valuable in the treatment of COVID-19. |
| publishDate |
2022 |
| dc.date.accessioned.none.fl_str_mv |
2022-06-03T15:15:20Z |
| dc.date.available.none.fl_str_mv |
2022-06-03T15:15:20Z |
| dc.date.issued.none.fl_str_mv |
2022-03-18 |
| dc.type.none.fl_str_mv |
Artículos Científicos |
| dc.type.coar.none.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| dc.type.coarversion.none.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.driver.none.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.redcol.none.fl_str_mv |
http://purl.org/redcol/resource_type/ART |
| dc.type.version.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| status_str |
publishedVersion |
| dc.identifier.uri.spa.fl_str_mv |
10.3389/fmicb.2022.721103. PMID: 35369500 |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/20.500.12494/45190 |
| dc.identifier.bibliographicCitation.spa.fl_str_mv |
Zapata-Cardona MI, Flórez-Álvarez L, Zapata-Builes W, Guerra-Sandoval AL, Guerra-Almonacid CM, Hincapié-García J, Rugeles MT, Hernandez JC. Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro. Front Microbiol. 2022 Mar 18;13:721103. doi: 10.3389/fmicb.2022.721103. PMID: 35369500; PMCID: PMC8972052. |
| identifier_str_mv |
10.3389/fmicb.2022.721103. PMID: 35369500 Zapata-Cardona MI, Flórez-Álvarez L, Zapata-Builes W, Guerra-Sandoval AL, Guerra-Almonacid CM, Hincapié-García J, Rugeles MT, Hernandez JC. Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro. Front Microbiol. 2022 Mar 18;13:721103. doi: 10.3389/fmicb.2022.721103. PMID: 35369500; PMCID: PMC8972052. |
| url |
https://hdl.handle.net/20.500.12494/45190 |
| dc.relation.ispartofjournal.spa.fl_str_mv |
Frontiers in Microbiology |
| dc.relation.references.spa.fl_str_mv |
Abdelnabi, R. (2020). Atorvastatin as Adjunctive Therapy in COVID-19 (STATCO19). National Library of Medicine: United States Abdelnabi, R., Foo, C. S., de Jonghe, S., Maes, P., Weynand, B., and Neyts, J. (2021). Molnupiravir inhibits replication of the emerging SARS-CoV-2 variants of concern in a hamster infection model. J. Infect. Dis. 224, 749–753. doi: 10.1093/infdis/jiab361 Acharya, A., Pandey, K., Thurman, M., Klug, E., Trivedi, J., Sharma, K., et al. (2021). Discovery and evaluation of entry inhibitors for SARS-CoV-2 and its emerging variants. J. Virol. 95, e01437–e01421. doi: 10.1128/ JVI.01437-21 Ader, F., Bouscambert-Duchamp, M., Hites, M., Peiffer-Smadja, N., Poissy, J., Belhadi, D., et al. (2021). Remdesivir plus standard of care versus standard of care alone for the treatment of patients admitted to hospital with COVID-19 (DisCoVeRy): a phase 3, randomised, controlled, open-label trial. Lancet Infect. Dis. 22, 209–221. doi: 10.1016/S1473-3099(21)00485-0 Ahmad, J., Ikram, S., Ahmad, F., Rehman, I. U., and Mushtaq, M. (2020). SARS-CoV-2 RNA dependent RNA polymerase (RdRp) – a drug repurposing study. Heliyon 6:e04502. doi: 10.1016/j.heliyon.2020.e04502 Alexpandi, R., de Mesquita, J. F., Pandian, S. K., and Ravi, A. V. (2020). Quinolines-based SARS-CoV-2 3CLpro and RdRp inhibitors and spike-RBDACE2 inhibitor for drug-repurposing Against COVID-19: an in silico analysis. Front. Microbiol. 11:1796. doi: 10.3389/fmicb.2020.01796 Al-Horani, R. A., Kar, S., and Aliter, K. F. (2020). Potential anti-COVID-19 therapeutics that block the early stage of the viral life cycle: structures, mechanisms, and clinical trials. Int. J. Mol. Sci. 21:5224. doi: 10.3390/ ijms21155224 Baby, K., et al. (2020). Targeting SARS-CoV-2 RNA-dependent RNA polymerase: an in silico drug repurposing for COVID-19. F1000Res. 9:1166. doi: 10.12688/ f1000research.26359.1 Bajimaya, S., Hayashi, T., Frankl, T., Bryk, P., Ward, B., and Takimoto, T. (2017). Cholesterol reducing agents inhibit assembly of type I parainfluenza viruses. Virology 501, 127–135. doi: 10.1016/j.virol.2016.11.011 Berman, H. M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T. N., Weissig, H., et al. (2000). The Protein Data Bank. Nucleic Acids Res. 28, 235–242. doi: 10.1093/nar/28.1.235 BIOVIA (2020), D.S. Discovery Studio Visualizer Software, Version 16.1 2017. Available at: https://discover.3ds.com/discovery-studio-visualizer-download [Aceesed December, 2020]. Björkhem-Bergman, L., Lindh, J. D., and Bergman, P. (2011). What is a relevant statin concentration in cell experiments claiming pleiotropic effects? Br. J. Clin. Pharmacol. 72, 164–165. doi: 10.1111/j.1365-2125.2011.03907.x Blaess, M., Kaiser, L., Sauer, M., Csuk, R., and Deigner, H.-P. (2020). COVID-19/ SARS-CoV-2 Infection: Lysosomes and Lysosomotropism Implicate New Treatment Strategies and Personal Risks. Int. J. Mol. Sci. 21:4953. doi: 10.3390/ ijms21144953 Boopathi, S., Poma, A. B., and Kolandaivel, P. (2020). Novel 2019 coronavirus structure, mechanism of action, antiviral drug promises and rule out against its treatment. J. Biomol. Struct. Dyn. 39, 1–10. doi: 10.1080/07391102.2020.1758788 Bryan-Marrugo, O. L., Arellanos-Soto, D., Rojas-Martinez, A., Barrera-Saldaña, H., Ramos-Jimenez, J., Vidaltamayo, R., et al. (2016). The anti-dengue virus properties of statins may be associated with alterations in the cellular antiviral profile expression. Mol. Med. Rep. 14, 2155–2163. doi: 10.3892/mmr.2016.5519 Calligari, P., Bobone, S., Ricci, G., and Bocedi, A. (2020). Molecular investigation of SARS-CoV-2 proteins and their interactions with antiviral drugs. Viruses 12:445. doi: 10.3390/v12040445 Castiglione, V., Chiriacò, M., Emdin, M., Taddei, S., and Vergaro, G. (2020). Statin therapy in COVID-19 infection. Eur. Heart J. 6, 258–259. doi: 10.1093/ ehjcvp/pvaa042 Choi, G. J., Kim, H. M., and Kang, H. (2020). The potential role of dyslipidemia in COVID-19 severity: an umbrella review of systematic reviews. J. Lipid Atheroscler. 9, 435–448. doi: 10.12997/jla.2020.9.3.435 Clausen, T. M., et al., (2020). SARS-CoV-2 infection depends on cellular heparan sulfate and ACE2. bioRxiv [Preprint]. ClinicalTrials.gov (2020). Intermediate-dose vs standard prophylactic anticoagulation and statin vs placebo in ICU patients with COVID-19 (INSPIRATION). National Library of Medicine: United States Conzelmann, C., Gilg, A., Groß, R., Schütz, D., Preising, N., Ständker, L., et al. (2020). An enzyme-based immunodetection assay to quantify SARSCoV-2 infection. Antivir. Res. 181:104882. doi: 10.1016/j.antiviral.2020.10488 Corman, V. M., Landt, O., Kaiser, M., Molenkamp, R., Meijer, A., Chu, D. K., et al. (2020). Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 25:2000045. doi: 10.2807/1560-7917.ES.2020.25.3.2000045 da Silva Gomes Dias, S., Soares, V. C., Ferreira, A. C., Sacramento, C. Q., Fintelman-Rodrigues, N., Temerozo, J. R., et al. (2020). Lipid droplets fuel SARS-CoV-2 replication and production of inflammatory mediators. PLoS Pathog. 16:e1009127. doi: 10.1371/journal.ppat.1009127 Delang, L., Paeshuyse, J., Vliegen, I., Leyssen, P., Obeid, S., Durantel, D., et al. (2009). Statins potentiate the in vitro anti-hepatitis C virus activity of selective hepatitis C virus inhibitors and delay or prevent resistance development. Hepatology 50, 6–16. doi: 10.1002/hep.22916 Devaux, C. A., Rolain, J. M., Colson, P., and Raoult, D. (2020). New insights on the antiviral effects of chloroquine against coronavirus: what to expect for COVID-19? Int. J. Antimicrob. Agents 55:105938. doi: 10.1016/j. ijantimicag.2020.105938 Díaz, F. J., Aguilar-Jiménez, W., Flórez-Álvarez, L., Valencia, G., Laiton-Donato, K., Franco-Muñoz, C., et al. (2020). Aislamiento y caracterización de una cepa temprana de SARS-CoV-2 durante la epidemia de 2020 en Medellín, Colombia. Biomédica 40, 148–158. doi: 10.7705/biomedica.5834 Ding, S., Yu, B., and van Vuuren, A. J. (2021). Statins significantly repress rotavirus replication through downregulation of cholesterol synthesis. Gut Microbes 13:1955643. doi: 10.1080/19490976.2021.1955643 Episcopio, D., Aminov, S., Benjamin, S., Germain, G., Datan, E., Landazuri, J., et al. (2019). Atorvastatin restricts the ability of influenza virus to generate lipid droplets and severely suppresses the replication of the virus. FASEB J. 33, 9516–9525. doi: 10.1096/fj.201900428RR Esakandari, H., Nabi-Afjadi, M., Fakkari-Afjadi, J., Farahmandian, N., Miresmaeili, S. M., and Bahreini, E. (2020). A comprehensive review of COVID-19 characteristics. Biol. Proced. Online 22:19. doi: 10.1186/ s12575-020-00128-2 Españo, E., Nam, J. H., Song, E. J., Song, D., Lee, C. K., and Kim, J. K. (2019). Lipophilic statins inhibit Zika virus production in Vero cells. Sci. Rep. 9, 11461–11461. doi: 10.1038/s41598-019-47956-1 FDA (2020). Coronavirus (COVID-19) Update: FDA Authorizes Drug Combination for Treatment of COVID-19. Available at: https://www.fda.gov/news-events/ press-announcements/coronavirus-covid-19-update-fda-authorizes-drugcombination-treatment-covid-19 (Accessed January, 2021) FDA (2021a). FDA Approves First Treatment for COVID-19. Available at: https://www.fda.gov/news-events/press-announcements/fda-approves-firsttreatment-covid-19 (Accessed December, 2021) FDA (2021b). Coronavirus (COVID-19) Update: FDA Authorizes Drug Combination for Treatment of COVID-19. Available at: https://www.fda. gov/news-events/press-announcements/coronavirus-covid-19-update-fdaauthorizes-drug-combination-treatment-covid-19 Fedson, D. S., Opal, S. M., and Rordam, O. M. (2020). Hiding in plain sight: an approach to treating patients with severe COVID-19 Infection. MBio 11, e00398–e00320. doi: 10.1128/mBio.00398-20 Feng, B., Xu, L., Wang, H., Yan, X., Xue, J., Liu, F., et al. (2011). Atorvastatin exerts its anti-atherosclerotic effects by targeting the receptor for advanced glycation end products. Biochim. Biophys. Acta 1812, 1130–1137. doi: 10.1016/j. bbadis.2011.05.007 Ganjali, S., Bianconi, V., Penson, P. E., Pirro, M., Banach, M., Watts, G. F., et al. (2020). Commentary: statins, COVID-19, and coronary artery disease: killing two birds with one stone. Metab. Clin. Exp. 113, 154375–154375. doi: 10.1016/j.metabol.2020.154375 Gao, Y., Yan, L., Huang, Y., Liu, F., Zhao, Y., Cao, L., et al. (2020). Structure of the RNA-dependent RNA polymerase from COVID-19 virus. Science 368, 779–782. doi: 10.1126/science.abb7498 González-Rayasa, J., Ana, R.-G., José, G.-G., José, G.-Y., José, H.-H., and, Rosa, del Carmen L.-S. (2020). COVID-19 and ACE -inhibitors and angiotensin receptor blockers-: The need to differentiate between early infection and acute lung injury. Rev. Colomb. de Cardiol. 27, 129–131. doi: 10.1016/j. rccar.2020.04.005 Greenwood, J., Steinman, L., and Zamvil, S. S. (2006). Statin therapy and autoimmune disease: from protein prenylation to immunomodulation. Nat. Rev. Immunol. 6, 358–370. doi: 10.1038/nri1839 Haidari, M., Ali, M., Casscells, S. W., and Madjid, M. (2007). Statins block influenza infection by down-regulating rho/rho kinase pathway. Circulation 116, 116–117. doi: 10.1161/circ.116.suppl_16.II_7 Hanwell, M. D., Curtis, D. E., Lonie, D. C., Vandermeersch, T., Zurek, E., and Hutchison, G. R. (2012). Avogadro: an advanced semantic chemical editor, visualization, and analysis platform. J. Chemother. 4:17. doi: 10.1186/1758-2946-4-17 Harisna, A. H., Nurdiansyah, R., Syaifie, P. H., Nugroho, D. W., Saputro, K. E., Firdayani, , et al. (2021). In silico investigation of potential inhibitors to main protease and spike protein of SARS-CoV-2 in propolis. Biochem. Biophys. Rep. 26:100969. doi: 10.1016/j.bbrep.2021.100969 Isusi, E., Aspichueta, P., Liza, M., Hernández, M.́. L., Dı́az, C., Hernández, G., et al. (2000). Short- and long-term effects of atorvastatin, lovastatin and simvastatin on the cellular metabolism of cholesteryl esters and VLDL secretion in rat hepatocytes. Atherosclerosis 153, 283–294. doi: 10.1016/ S0021-9150(00)00407-X Jendele, L., Krivak, R., Skoda, P., Novotny, M., and Hoksza, D. (2019). PrankWeb: a web server for ligand binding site prediction and visualization. Nucleic Acids Res. 47, W345–W349. doi: 10.1093/nar/gkz424 Kalil, A. C., Patterson, T. F., Mehta, A. K., Tomashek, K. M., Wolfe, C. R., Ghazaryan, V., et al. (2020). Baricitinib plus Remdesivir for hospitalized adults with Covid-19. N. Engl. J. Med. 384, 795–807. doi: 10.1056/NEJMoa2031994 Khateeb, J., Li, Y., and Zhang, H. (2021). Emerging SARS-CoV-2 variants of concern and potential intervention approaches. Crit. Care 25, 244–244. doi: 10.1186/s13054-021-03662-x Kim, S. S., Peng, L. F., Lin, W., Choe, W.-H., Sakamoto, N., Kato, N., et al. (2007). A cell-based, high-throughput screen for small molecule regulators of hepatitis C virus replication. Gastroenterology 132, 311–320. doi: 10.1053/j. gastro.2006.10.032 Klein, S., Cortese, M., Winter, S. L., Wachsmuth-Melm, M., Neufeldt, C. J., Cerikan, B., et al. (2020). SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography. Nat. Commun. 11:5885. doi: 10.1038/ s41467-020-19619-7 Korber, B., Fischer, W. M., Gnanakaran, S., Yoon, H., Theiler, J., Abfalterer, W., et al. (2020). Tracking changes in SARS-CoV-2 spike: evidence that D614G increases infectivity of the COVID-19 virus. Cell 182, 812–827.e19. doi: 10.1016/j.cell.2020.06.043 Koulgi, S., Jani, V., Uppuladinne, M. V. N., Sonavane, U., and Joshi, R. (2020). Remdesivir-bound and ligand-free simulations reveal the probable mechanism of inhibiting the RNA dependent RNA polymerase of severe acute respiratory syndrome coronavirus 2. RSC Adv. 10, 26792–26803. doi: 10.1039/D0RA04743K Kow, C. S., and Hasan, S. S. (2020). Meta-analysis of effect of statins in patients with COVID-19. Am. J. Cardiol. 134, 153–155. doi: 10.1016/j. amjcard.2020.08.004 Kumar, R., Mehta, D., Mishra, N., Nayak, D., and Sunil, S. (2020). Role of host-mediated post-translational modifications (PTMs) in RNA virus pathogenesis. Int. J. Mol. Sci. 22:323. doi: 10.3390/ijms22010323 Kumar, Y., Singh, H., and Patel, C. N. (2020). In silico prediction of potential inhibitors for the main protease of SARS-CoV-2 using molecular docking and dynamics simulation based drug-repurposing. J. Infect. Public Health 13, 1210–1223. doi: 10.1016/j.jiph.2020.06.016 Kwon, P. S., Oh, H., Kwon, S. J., Jin, W., Zhang, F., Fraser, K., et al. (2020). Sulfated polysaccharides effectively inhibit SARS-CoV-2 in vitro. Cell Dis. 6:50. doi: 10.1038/s41421-020-00192-8 Li, Z., Li, X., Huang, Y. Y., Wu, Y., Liu, R., Zhou, L., et al. (2020). Identify potent SARS-CoV-2 main protease inhibitors via accelerated free energy perturbation-based virtual screening of existing drugs. Proc. Natl. Acad. Sci. 117, 27381–27387. doi: 10.1073/pnas.2010470117 Lotfi, M., Hamblin, M. R., and Rezaei, N. (2020). COVID-19: transmission, prevention, and potential therapeutic opportunities. Clin. Chim. Acta 508, 254–266. doi: 10.1016/j.cca.2020.05.044 Lu, Y., Liu, D. X., and Tam, J. P. (2008). Lipid rafts are involved in SARS-CoV entry into Vero E6 cells. Biochem. Biophys. Res. Commun. 369, 344–349. doi: 10.1016/j.bbrc.2008.02.023 Marín-Palma, D., Tabares-Guevara, J. H., Zapata-Cardona, M. I., Flórez-Álvarez, L., Yepes, L. M., Rugeles, M. T., et al. (2021). Curcumin inhibits in vitro SARS-CoV-2 infection in vero E6 cells through multiple antiviral mechanisms. Molecules 26:6900. doi: 10.3390/molecules26226900 Martínez-Gutierrez, M., Castellanos, J. E., and Gallego-Gómez, J. C. (2011). Statins reduce dengue virus production via decreased virion assembly. Intervirology 54, 202–216. doi: 10.1159/000321892 Matsushita, K., Ding, N., Kou, M., Hu, X., Chen, M., Gao, Y., et al. (2020). The relationship of COVID-19 severity with cardiovascular disease and its traditional risk factors: a systematic review and meta-analysis. Glob. Heart 15, 64–64. doi: 10.5334/gh.814 Mehrbod, P., Ideris, A., Omar, A. R., and Hair-Bejo, M. (2012). Evaluation of antiviral effect of atorvastatin on H1N1 infection in MDCK cells. Afr. J. Microbiol. Res. 6, 5715–5719. doi: 10.5897/AJMR12.1011 Mendonça, L., Howe, A, Gilchrist, JB, Sun, D, Knight, ML, Zanetti-Domingues, LC, et al., (2020). SARS-CoV-2 assembly and egress pathway revealed by correlative multi-modal multi-scale Cryo-imaging. bioRxiv [Preprint]. Mendoza, E. J., Manguiat, K., Wood, H., and Drebot, M. (2020). Two Detailed Plaque Assay Protocols for the Quantification of Infectious SARS-CoV-2. Curr. Protoc. Microbiol. 57:ecpmc105. doi: 10.1002/cpmc.105 Minz, M. M., Bansal, M., and Kasliwal, R. R. (2020). Statins and SARS-CoV-2 disease: current concepts and possible benefits. Diabetes Metab. Syndr. 14, 2063–2067. doi: 10.1016/j.dsx.2020.10.021 Mukherjee, S., Bhattacharyya, D., and Bhunia, A. (2020). Host-membrane interacting interface of the SARS coronavirus envelope protein: immense functional potential of C-terminal domain. Biophys. Chem. 266, 106452–106452. doi: 10.1016/j.bpc.2020.106452 Mycroft-West, C. J., Su, D., Pagani, I., Rudd, T. R., Elli, S., Gandhi, N. S., et al. (2020). Heparin inhibits cellular invasion by SARS-CoV-2: structural dependence of the interaction of the spike S1 receptor-binding domain with heparin. Thromb. Haemost. 120, 1700–1715. doi: 10.1055/s-0040- 1721319 Naik, V. R., Munikumar, M., Ramakrishna, U., Srujana, M., Goudar, G., Naresh, P., et al. (2020). Remdesivir (GS-5734) as a therapeutic option of 2019-nCOV main protease - in silico approach. J. Biomol. Struct. Dyn. 39, 4701–4714. doi: 10.1080/07391102.2020.1781694 Nimgampalle, M., Devanathan, V., and Saxena, A. (2020). Screening of Chloroquine, Hydroxychloroquine and its derivatives for their binding affinity to multiple SARS-CoV-2 protein drug targets. J. Biomol. Struct. Dyn. 39, 4949–4961. doi: 10.1080/07391102.2020.1782265 Otto, S. P., Day, T., Arino, J., Colijn, C., Dushoff, J., Li, M., et al. (2021). The origins and potential future of SARS-CoV-2 variants of concern in the evolving COVID-19 pandemic. Curr. Biol. 31, R918–R929. doi: 10.1016/j. cub.2021.06.049 https://doi.org/10.3389/fmolb.2020.578964 Pan, H., Peto, R., Henao-Restrepo, A. M., Preziosi, M. P., Sathiyamoorthy, V., Abdool Karim, Q., et al. (2021). Repurposed antiviral drugs for Covid-19 - interim WHO solidarity trial results. N. Engl. J. Med. 384, 497–511. doi: 10.1056/NEJMoa2023184 Parquet, V., Henry, M., Wurtz, N., Dormoi, J., Briolant, S., Gil, M., et al. (2010). Atorvastatin as a potential anti-malarial drug: in vitro synergy in combinational therapy with quinine against plasmodium falciparum. Malar. J. 9:139. doi: 10.1186/1475-2875-9-139 Pawlos, A., Niedzielski, M., Gorzelak-Pabiś, P., Broncel, M., and Woźniak, E. (2021). COVID-19: direct and indirect mechanisms of statins. Int. J. Mol. Sci. 22, 4177. doi: 10.3390/ijms22084177 PFIZER (2010). Pfizer announces European Union approval of a new form of lipitor (atorvastatin) for use in children. Available at: https://www.pfizer. com/news/press-release/press-release-detail/pfizer_announces_european_ union_approval_of_a_new_form_of_lipitor_atorvastatin_for_use_in_children (Accessed January, 2021). Reiner, Ž., Hatamipour, M., Banach, M., Pirro, M., al-Rasadi, K., Jamialahmadi, T., et al. (2020). Statins and the COVID-19 main protease: in silico evidence on direct interaction. Arch. Med. Sci. 16, 490–496. doi: 10.5114/aoms.2020.94655 Rudrapal, M., Khairnar, S., and Jadhav, A. (2020). Drug Repurposing (DR): An Emerging Approach in Drug Discovery. London: IntechOpen. Sabatino, J., de Rosa, S., di Salvo, G., and Indolfi, C. (2020). Impact of cardiovascular risk profile on COVID-19 outcome. A meta-analysis. PLoS One 15:e0237131. doi: 10.1371/journal.pone.0237131 Sacco, M. D., Ma, C., Lagarias, P., Gao, A., Townsend, J. A., Meng, X., et al. (2020). Structure and inhibition of the SARS-CoV-2 main protease reveal strategy for developing dual inhibitors against Mspro and cathepsin L. Sci. Adv. 6:eabe0751. doi: 10.1126/sciadv.abe0751 Sanders, D. W., et al., (2020). SARS-CoV-2 requires cholesterol for viral entry and pathological syncytia formation. bioRxiv [preprint]. Schmidt, N. M., Wing, P. A. C., McKeating, J. A., and Maini, M. K. (2020). Cholesterol-modifying drugs in COVID-19. Oxf. open immunol. 1:iqaa001. doi: 10.1093/oxfimm/iqaa001 Schöning-Stierand, K., Diedrich, K., Fährrolfes, R., Flachsenberg, F., Meyder, A., Nittinger, E., et al. (2020). ProteinsPlus: interactive analysis of protein–ligand binding interfaces. Nucleic Acids Res. 48, W48–W53. doi: 10.1093/nar/gkaa235 Shrivastava-Ranjan, P., Flint, M., Bergeron, É., McElroy, A. K., Chatterjee, P., Albariño, C. G., et al. (2018). Statins Suppress Ebola Virus Infectivity by Interfering with Glycoprotein Processing. MBio 9, e00660–e00618. doi: 10.1128/mBio.00660-18 Simabuco, F. M., Tamura, R. E., Pavan, I. C. B., Morale, M. G., and Ventura, A. M. (2021). Molecular mechanisms and pharmacological interventions in the replication cycle of human coronaviruses. Genet. Mol. Biol. 44:e20200212. doi: 10.1590/1678-4685-gmb-2020-0212 Singh, D. D., Han, I., Choi, E. H., and Yadav, D. K. (2020). Recent advances in pathophysiology, drug development and future perspectives of SARSCoV-2. Front. Cell Dev. Biol. 8:580202. doi: 10.3389/fcell.2020.580202 Su, H. X., Yao, S., Zhao, W. F., Li, M. J., Liu, J., Shang, W. J., et al. (2020). Anti-SARS-CoV-2 activities in vitro of Shuanghuanglian preparations and bioactive ingredients. Acta Pharmacol. Sin. 41, 1167–1177. doi: 10.1038/ s41401-020-0483-6 Su, H. X., Yao, S., Zhao, W. F., Li, M. J., Liu, J., Shang, W. J., et al. (2020). Anti-SARS-CoV-2 activities in vitro of Shuanghuanglian preparations and bioactive ingredients. Acta Pharmacol. Sin. 41, 1167–1177. doi: 10.1038/ s41401-020-0483-6 Tandon, R., Sharp, J. S., Zhang, F., Pomin, V. H., Ashpole, N. M., Mitra, D., et al. (2021). Effective inhibition of SARS-CoV-2 entry by heparin and enoxaparin derivatives. J. Virol. 95, e01987–e01920. doi: 10.1128/ JVI.01987-20 Trott, O., and Olson, A. J. (2010). AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 31, 455–461. doi: 10.1002/jcc.21334 Uzunova, K., Filipova, E., Pavlova, V., and Vekov, T. (2020). Insights into antiviral mechanisms of remdesivir, lopinavir/ritonavir and chloroquine/ hydroxychloroquine affecting the new SARS-CoV-2. Biomed. Pharmacother. 131:110668. doi: 10.1016/j.biopha.2020.110668 TUzunova, K., Filipova, E., Pavlova, V., and Vekov, T. (2020). Insights into antiviral mechanisms of remdesivir, lopinavir/ritonavir and chloroquine/ hydroxychloroquine affecting the new SARS-CoV-2. Biomed. Pharmacother. 131:110668. doi: 10.1016/j.biopha.2020.110668 Villareal, V. A., Rodgers, M. A., Costello, D. A., and Yang, P. L. (2015). Targeting host lipid synthesis and metabolism to inhibit dengue and hepatitis C viruses. Antivir. Res. 124, 110–121. doi: 10.1016/j.antiviral.2015.10.013 V’kovski, P., Kratzel, A., Steiner, S., Stalder, H., and Thiel, V. (2021). Coronavirus biology and replication: implications for SARS-CoV-2. Biomed. Pharmacother. 19, 155–121. doi: 10.1038/s41579-020-00468-6 Wang, M., Cao, R., Zhang, L., Yang, X., Liu, J., X Wang, H., Yuan, Z, Pavel, M. A., Jablonski, S. M., Jablonski, J., Hobson, R., et al., (2021). The role of high cholesterol in age-related COVID19 lethality. bioRxiv [Preprint]. doi: 10.1101/2020.05.09.086249 Wang, M., Cao, R., Zhang, L., Yang, X., Liu, J., Xu, M., et al. (2020). Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro. Cell Res. 30, 269–271. doi: 10.1038/ s41422-020-0282-0 Wang, Y., Zhang, D., du, G., du, R., Zhao, J., Jin, Y., et al. (2020). Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial. Lancet 395, 1569–1578. doi: 10.1016/S0140-6736(20) 31022-9 WHO (2020a). COVID-19: cronología de la actuación de la OMS. Available at: https://www.who.int/es/news/item/27-04-2020-who-timeline---covid-19 (Accessed January, 2022). WHO (2020b). WHO Coronavirus Disease (COVID-19) Dashboard. Available at: https://covid19.who.int/ (Accessed January, 2021). Wolff, G., Melia, C. E., Snijder, E. J., and Bárcena, M. (2020). Double-membrane vesicles as platforms for viral replication. Trends Microbiol. 28, 1022–1033. doi: 10.1016/j.tim.2020.05.009 Wösten-van Asperen, R. M., Bos, A. P., Bem, R. A., Dierdorp, B. S., Dekker, T., van Goor, H., et al. (2013). Imbalance between pulmonary angiotensinconverting enzyme and angiotensin-converting enzyme 2 activity in acute respiratory distress syndrome. Pediatr. Crit. Care Med. 14, e438–e441. doi: 10.1097/PCC.0b013e3182a55735 Wurtz, N., Penant, G., Jardot, P., Duclos, N., and la Scola, B. (2021). Culture of SARS-CoV-2 in a panel of laboratory cell lines, permissivity, and differences in growth profile. EJCDEU 40, 477–484. doi: 10.1007/ s10096-020-04106-0 Yepes-Perez, A. F., Herrera-Calderón, O., Oliveros, C. A., Flórez-Álvarez, L., Zapata-Cardona, M. I., Yepes, L., et al. (2021). The Hydroalcoholic extract Yuan, S., Chan, C. C. Y., Chik, K. K. H., Tsang, J. O. L., Liang, R., Cao, J., et al. (2020). Broad-Spectrum host-based antivirals targeting the interferon and lipogenesis pathways as potential treatment options for the pandemic coronavirus disease 2019 (COVID-19). Viruses 12:628. doi: 10.3390/ v12060628 Zapatero-Belinchón, F. J., et al. (2021). Fluvastatin mitigates SARS-CoV-2 infection in human lung cells. iScience 24:103469. doi: 10.1016/j.isci.2021. 103469 Zhang, Q., Chen, C. Z., Swaroop, M., Xu, M., Wang, L., Lee, J., et al. (2020). Heparan sulfate assists SARS-CoV-2 in cell entry and can be targeted by approved drugs in vitro. Cell Dis. 6:80. doi: 10.1038/s41421-020-00222-5 |
| dc.rights.license.none.fl_str_mv |
Atribución |
| dc.rights.accessrights.none.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.coar.none.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| rights_invalid_str_mv |
Atribución http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.coverage.temporal.spa.fl_str_mv |
13 |
| dc.publisher.spa.fl_str_mv |
Universidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Programa de Medicina, Medellín y Envigado, Colombia, 00000 |
| dc.publisher.program.spa.fl_str_mv |
Medicina |
| dc.publisher.place.spa.fl_str_mv |
Medellín |
| institution |
Universidad Cooperativa de Colombia |
| bitstream.url.fl_str_mv |
https://repository.ucc.edu.co/bitstreams/f7441486-babe-4131-890c-366a8f9e3998/download https://repository.ucc.edu.co/bitstreams/648d10dc-ed05-4306-a8f0-67d0f2b999a4/download https://repository.ucc.edu.co/bitstreams/afbd3188-ac71-4bc4-b45c-fb5b51935df2/download https://repository.ucc.edu.co/bitstreams/0b94634a-975f-48e4-add7-12f7c942b97f/download |
| bitstream.checksum.fl_str_mv |
93c6a2a9cb98625ec4fa1b31c88155a0 8a4605be74aa9ea9d79846c1fba20a33 e171505baddf792665a539622779d725 59a33f910fb8fc94513c2e4ff101532b |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Institucional Universidad Cooperativa de Colombia |
| repository.mail.fl_str_mv |
bdigital@metabiblioteca.com |
| _version_ |
1811565079861133312 |
| spelling |
Zapata-Cardona, María IFlórez-Álvarez, LizdanyHernández López, Juan Carlos Zapata Builes, WildemanGuerra-Sandoval, Ariadna LHincapié-García, JaimeRugeles, Maria T132022-06-03T15:15:20Z2022-06-03T15:15:20Z2022-03-1810.3389/fmicb.2022.721103. PMID: 35369500https://hdl.handle.net/20.500.12494/45190Zapata-Cardona MI, Flórez-Álvarez L, Zapata-Builes W, Guerra-Sandoval AL, Guerra-Almonacid CM, Hincapié-García J, Rugeles MT, Hernandez JC. Atorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitro. Front Microbiol. 2022 Mar 18;13:721103. doi: 10.3389/fmicb.2022.721103. PMID: 35369500; PMCID: PMC8972052.This article evaluated the in vitro antiviral effect of atorvastatin (ATV) against SARS-CoV-2 and identified the interaction affinity between this compound and two SARS-CoV-2 proteins. The antiviral activity of atorvastatin against this virus was evaluated by three different treatment strategies [(i) pre-post treatment, (ii) pre-infection treatment, and (iii) post-infection treatment] using Vero E6 and Caco-2 cells. The interaction of atorvastatin with RdRp (RNA-dependent RNA polymerase) and 3CL protease (3-chymotrypsin-like protease) was evaluated by molecular docking. The CC50s (half-maximal cytotoxic concentrations) obtained for ATV were 50.3 and 64.5 μM in Vero E6 and Caco-2, respectively. This compound showed antiviral activity against SARS-CoV-2 D614G strain in Vero E6 with median effective concentrations (EC50s) of 15.4, 12.1, and 11.1μM by pre-post, pre-infection, and post-infection treatments, respectively. ATV also inhibited Delta and Mu variants by pre-post treatment (EC50s of 16.8 and 21.1μM, respectively). In addition, ATV showed an antiviral effect against the D614G strain independent of the cell line (EC50 of 7.4μM in Caco-2). The interaction of atorvastatin with SARS-CoV-2 RdRp and 3CL protease yielded a binding affinity of −6.7kcal/mol and −7.5kcal/mol, respectively. Our study demonstrated the in vitro antiviral activity of atorvastatin against the ancestral SARS-CoV-2 D614G strain and two emerging variants (Delta and Mu), with an independent effect of the cell line. A favorable binding affinity between ATV and viral proteins by bioinformatics methods was found. Due to the extensive clinical experience of atorvastatin use, it could prove valuable in the treatment of COVID-19.https://scienti.minciencias.gov.co/cvlac/visualizador/generarCurriculoCv.do?cod_rh=0000283088http://orcid.org/0000-0002-9200-5698https://scienti.colciencias.gov.co/gruplac/jsp/visualiza/visualizagr.jsp?nro=00000000011355juankhernandez@gmail.comUniversidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Programa de Medicina, Medellín y Envigado, Colombia, 00000MedicinaMedellínSARS-CoV-2antiviralmolecular dockingCOVID-19variantsAtorvastatin Effectively Inhibits Ancestral and Two Emerging Variants of SARS-CoV-2 in vitroArtículos Científicoshttp://purl.org/coar/resource_type/c_2df8fbb1http://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionAtribucióninfo:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Frontiers in MicrobiologyAbdelnabi, R. (2020). Atorvastatin as Adjunctive Therapy in COVID-19 (STATCO19). National Library of Medicine: United StatesAbdelnabi, R., Foo, C. S., de Jonghe, S., Maes, P., Weynand, B., and Neyts, J. (2021). Molnupiravir inhibits replication of the emerging SARS-CoV-2 variants of concern in a hamster infection model. J. Infect. Dis. 224, 749–753. doi: 10.1093/infdis/jiab361Acharya, A., Pandey, K., Thurman, M., Klug, E., Trivedi, J., Sharma, K., et al. (2021). Discovery and evaluation of entry inhibitors for SARS-CoV-2 and its emerging variants. J. Virol. 95, e01437–e01421. doi: 10.1128/ JVI.01437-21Ader, F., Bouscambert-Duchamp, M., Hites, M., Peiffer-Smadja, N., Poissy, J., Belhadi, D., et al. (2021). Remdesivir plus standard of care versus standard of care alone for the treatment of patients admitted to hospital with COVID-19 (DisCoVeRy): a phase 3, randomised, controlled, open-label trial. Lancet Infect. Dis. 22, 209–221. doi: 10.1016/S1473-3099(21)00485-0Ahmad, J., Ikram, S., Ahmad, F., Rehman, I. U., and Mushtaq, M. (2020). SARS-CoV-2 RNA dependent RNA polymerase (RdRp) – a drug repurposing study. Heliyon 6:e04502. doi: 10.1016/j.heliyon.2020.e04502Alexpandi, R., de Mesquita, J. F., Pandian, S. K., and Ravi, A. V. (2020). Quinolines-based SARS-CoV-2 3CLpro and RdRp inhibitors and spike-RBDACE2 inhibitor for drug-repurposing Against COVID-19: an in silico analysis. Front. Microbiol. 11:1796. doi: 10.3389/fmicb.2020.01796Al-Horani, R. A., Kar, S., and Aliter, K. F. (2020). Potential anti-COVID-19 therapeutics that block the early stage of the viral life cycle: structures, mechanisms, and clinical trials. Int. J. Mol. Sci. 21:5224. doi: 10.3390/ ijms21155224Baby, K., et al. (2020). Targeting SARS-CoV-2 RNA-dependent RNA polymerase: an in silico drug repurposing for COVID-19. F1000Res. 9:1166. doi: 10.12688/ f1000research.26359.1Bajimaya, S., Hayashi, T., Frankl, T., Bryk, P., Ward, B., and Takimoto, T. (2017). Cholesterol reducing agents inhibit assembly of type I parainfluenza viruses. Virology 501, 127–135. doi: 10.1016/j.virol.2016.11.011Berman, H. M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T. N., Weissig, H., et al. (2000). The Protein Data Bank. Nucleic Acids Res. 28, 235–242. doi: 10.1093/nar/28.1.235BIOVIA (2020), D.S. Discovery Studio Visualizer Software, Version 16.1 2017. Available at: https://discover.3ds.com/discovery-studio-visualizer-download [Aceesed December, 2020].Björkhem-Bergman, L., Lindh, J. D., and Bergman, P. (2011). What is a relevant statin concentration in cell experiments claiming pleiotropic effects? Br. J. Clin. Pharmacol. 72, 164–165. doi: 10.1111/j.1365-2125.2011.03907.xBlaess, M., Kaiser, L., Sauer, M., Csuk, R., and Deigner, H.-P. (2020). COVID-19/ SARS-CoV-2 Infection: Lysosomes and Lysosomotropism Implicate New Treatment Strategies and Personal Risks. Int. J. Mol. Sci. 21:4953. doi: 10.3390/ ijms21144953Boopathi, S., Poma, A. B., and Kolandaivel, P. (2020). Novel 2019 coronavirus structure, mechanism of action, antiviral drug promises and rule out against its treatment. J. Biomol. Struct. Dyn. 39, 1–10. doi: 10.1080/07391102.2020.1758788Bryan-Marrugo, O. L., Arellanos-Soto, D., Rojas-Martinez, A., Barrera-Saldaña, H., Ramos-Jimenez, J., Vidaltamayo, R., et al. (2016). The anti-dengue virus properties of statins may be associated with alterations in the cellular antiviral profile expression. Mol. Med. Rep. 14, 2155–2163. doi: 10.3892/mmr.2016.5519Calligari, P., Bobone, S., Ricci, G., and Bocedi, A. (2020). Molecular investigation of SARS-CoV-2 proteins and their interactions with antiviral drugs. Viruses 12:445. doi: 10.3390/v12040445Castiglione, V., Chiriacò, M., Emdin, M., Taddei, S., and Vergaro, G. (2020). Statin therapy in COVID-19 infection. Eur. Heart J. 6, 258–259. doi: 10.1093/ ehjcvp/pvaa042Choi, G. J., Kim, H. M., and Kang, H. (2020). The potential role of dyslipidemia in COVID-19 severity: an umbrella review of systematic reviews. J. Lipid Atheroscler. 9, 435–448. doi: 10.12997/jla.2020.9.3.435Clausen, T. M., et al., (2020). SARS-CoV-2 infection depends on cellular heparan sulfate and ACE2. bioRxiv [Preprint].ClinicalTrials.gov (2020). Intermediate-dose vs standard prophylactic anticoagulation and statin vs placebo in ICU patients with COVID-19 (INSPIRATION). National Library of Medicine: United StatesConzelmann, C., Gilg, A., Groß, R., Schütz, D., Preising, N., Ständker, L., et al. (2020). An enzyme-based immunodetection assay to quantify SARSCoV-2 infection. Antivir. Res. 181:104882. doi: 10.1016/j.antiviral.2020.10488Corman, V. M., Landt, O., Kaiser, M., Molenkamp, R., Meijer, A., Chu, D. K., et al. (2020). Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 25:2000045. doi: 10.2807/1560-7917.ES.2020.25.3.2000045da Silva Gomes Dias, S., Soares, V. C., Ferreira, A. C., Sacramento, C. Q., Fintelman-Rodrigues, N., Temerozo, J. R., et al. (2020). Lipid droplets fuel SARS-CoV-2 replication and production of inflammatory mediators. PLoS Pathog. 16:e1009127. doi: 10.1371/journal.ppat.1009127Delang, L., Paeshuyse, J., Vliegen, I., Leyssen, P., Obeid, S., Durantel, D., et al. (2009). Statins potentiate the in vitro anti-hepatitis C virus activity of selective hepatitis C virus inhibitors and delay or prevent resistance development. Hepatology 50, 6–16. doi: 10.1002/hep.22916Devaux, C. A., Rolain, J. M., Colson, P., and Raoult, D. (2020). New insights on the antiviral effects of chloroquine against coronavirus: what to expect for COVID-19? Int. J. Antimicrob. Agents 55:105938. doi: 10.1016/j. ijantimicag.2020.105938Díaz, F. J., Aguilar-Jiménez, W., Flórez-Álvarez, L., Valencia, G., Laiton-Donato, K., Franco-Muñoz, C., et al. (2020). Aislamiento y caracterización de una cepa temprana de SARS-CoV-2 durante la epidemia de 2020 en Medellín, Colombia. Biomédica 40, 148–158. doi: 10.7705/biomedica.5834Ding, S., Yu, B., and van Vuuren, A. J. (2021). Statins significantly repress rotavirus replication through downregulation of cholesterol synthesis. Gut Microbes 13:1955643. doi: 10.1080/19490976.2021.1955643Episcopio, D., Aminov, S., Benjamin, S., Germain, G., Datan, E., Landazuri, J., et al. (2019). Atorvastatin restricts the ability of influenza virus to generate lipid droplets and severely suppresses the replication of the virus. FASEB J. 33, 9516–9525. doi: 10.1096/fj.201900428RREsakandari, H., Nabi-Afjadi, M., Fakkari-Afjadi, J., Farahmandian, N., Miresmaeili, S. M., and Bahreini, E. (2020). A comprehensive review of COVID-19 characteristics. Biol. Proced. Online 22:19. doi: 10.1186/ s12575-020-00128-2Españo, E., Nam, J. H., Song, E. J., Song, D., Lee, C. K., and Kim, J. K. (2019). Lipophilic statins inhibit Zika virus production in Vero cells. Sci. Rep. 9, 11461–11461. doi: 10.1038/s41598-019-47956-1FDA (2020). Coronavirus (COVID-19) Update: FDA Authorizes Drug Combination for Treatment of COVID-19. Available at: https://www.fda.gov/news-events/ press-announcements/coronavirus-covid-19-update-fda-authorizes-drugcombination-treatment-covid-19 (Accessed January, 2021)FDA (2021a). FDA Approves First Treatment for COVID-19. Available at: https://www.fda.gov/news-events/press-announcements/fda-approves-firsttreatment-covid-19 (Accessed December, 2021)FDA (2021b). Coronavirus (COVID-19) Update: FDA Authorizes Drug Combination for Treatment of COVID-19. Available at: https://www.fda. gov/news-events/press-announcements/coronavirus-covid-19-update-fdaauthorizes-drug-combination-treatment-covid-19Fedson, D. S., Opal, S. M., and Rordam, O. M. (2020). Hiding in plain sight: an approach to treating patients with severe COVID-19 Infection. MBio 11, e00398–e00320. doi: 10.1128/mBio.00398-20Feng, B., Xu, L., Wang, H., Yan, X., Xue, J., Liu, F., et al. (2011). Atorvastatin exerts its anti-atherosclerotic effects by targeting the receptor for advanced glycation end products. Biochim. Biophys. Acta 1812, 1130–1137. doi: 10.1016/j. bbadis.2011.05.007Ganjali, S., Bianconi, V., Penson, P. E., Pirro, M., Banach, M., Watts, G. F., et al. (2020). Commentary: statins, COVID-19, and coronary artery disease: killing two birds with one stone. Metab. Clin. Exp. 113, 154375–154375. doi: 10.1016/j.metabol.2020.154375Gao, Y., Yan, L., Huang, Y., Liu, F., Zhao, Y., Cao, L., et al. (2020). Structure of the RNA-dependent RNA polymerase from COVID-19 virus. Science 368, 779–782. doi: 10.1126/science.abb7498González-Rayasa, J., Ana, R.-G., José, G.-G., José, G.-Y., José, H.-H., and, Rosa, del Carmen L.-S. (2020). COVID-19 and ACE -inhibitors and angiotensin receptor blockers-: The need to differentiate between early infection and acute lung injury. Rev. Colomb. de Cardiol. 27, 129–131. doi: 10.1016/j. rccar.2020.04.005Greenwood, J., Steinman, L., and Zamvil, S. S. (2006). Statin therapy and autoimmune disease: from protein prenylation to immunomodulation. Nat. Rev. Immunol. 6, 358–370. doi: 10.1038/nri1839Haidari, M., Ali, M., Casscells, S. W., and Madjid, M. (2007). Statins block influenza infection by down-regulating rho/rho kinase pathway. Circulation 116, 116–117. doi: 10.1161/circ.116.suppl_16.II_7Hanwell, M. D., Curtis, D. E., Lonie, D. C., Vandermeersch, T., Zurek, E., and Hutchison, G. R. (2012). Avogadro: an advanced semantic chemical editor, visualization, and analysis platform. J. Chemother. 4:17. doi: 10.1186/1758-2946-4-17Harisna, A. H., Nurdiansyah, R., Syaifie, P. H., Nugroho, D. W., Saputro, K. E., Firdayani, , et al. (2021). In silico investigation of potential inhibitors to main protease and spike protein of SARS-CoV-2 in propolis. Biochem. Biophys. Rep. 26:100969. doi: 10.1016/j.bbrep.2021.100969Isusi, E., Aspichueta, P., Liza, M., Hernández, M.́. L., Dı́az, C., Hernández, G., et al. (2000). Short- and long-term effects of atorvastatin, lovastatin and simvastatin on the cellular metabolism of cholesteryl esters and VLDL secretion in rat hepatocytes. Atherosclerosis 153, 283–294. doi: 10.1016/ S0021-9150(00)00407-XJendele, L., Krivak, R., Skoda, P., Novotny, M., and Hoksza, D. (2019). PrankWeb: a web server for ligand binding site prediction and visualization. Nucleic Acids Res. 47, W345–W349. doi: 10.1093/nar/gkz424Kalil, A. C., Patterson, T. F., Mehta, A. K., Tomashek, K. M., Wolfe, C. R., Ghazaryan, V., et al. (2020). Baricitinib plus Remdesivir for hospitalized adults with Covid-19. N. Engl. J. Med. 384, 795–807. doi: 10.1056/NEJMoa2031994Khateeb, J., Li, Y., and Zhang, H. (2021). Emerging SARS-CoV-2 variants of concern and potential intervention approaches. Crit. Care 25, 244–244. doi: 10.1186/s13054-021-03662-xKim, S. S., Peng, L. F., Lin, W., Choe, W.-H., Sakamoto, N., Kato, N., et al. (2007). A cell-based, high-throughput screen for small molecule regulators of hepatitis C virus replication. Gastroenterology 132, 311–320. doi: 10.1053/j. gastro.2006.10.032Klein, S., Cortese, M., Winter, S. L., Wachsmuth-Melm, M., Neufeldt, C. J., Cerikan, B., et al. (2020). SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography. Nat. Commun. 11:5885. doi: 10.1038/ s41467-020-19619-7Korber, B., Fischer, W. M., Gnanakaran, S., Yoon, H., Theiler, J., Abfalterer, W., et al. (2020). Tracking changes in SARS-CoV-2 spike: evidence that D614G increases infectivity of the COVID-19 virus. Cell 182, 812–827.e19. doi: 10.1016/j.cell.2020.06.043Koulgi, S., Jani, V., Uppuladinne, M. V. N., Sonavane, U., and Joshi, R. (2020). Remdesivir-bound and ligand-free simulations reveal the probable mechanism of inhibiting the RNA dependent RNA polymerase of severe acute respiratory syndrome coronavirus 2. RSC Adv. 10, 26792–26803. doi: 10.1039/D0RA04743KKow, C. S., and Hasan, S. S. (2020). Meta-analysis of effect of statins in patients with COVID-19. Am. J. Cardiol. 134, 153–155. doi: 10.1016/j. amjcard.2020.08.004Kumar, R., Mehta, D., Mishra, N., Nayak, D., and Sunil, S. (2020). Role of host-mediated post-translational modifications (PTMs) in RNA virus pathogenesis. Int. J. Mol. Sci. 22:323. doi: 10.3390/ijms22010323Kumar, Y., Singh, H., and Patel, C. N. (2020). In silico prediction of potential inhibitors for the main protease of SARS-CoV-2 using molecular docking and dynamics simulation based drug-repurposing. J. Infect. Public Health 13, 1210–1223. doi: 10.1016/j.jiph.2020.06.016Kwon, P. S., Oh, H., Kwon, S. J., Jin, W., Zhang, F., Fraser, K., et al. (2020). Sulfated polysaccharides effectively inhibit SARS-CoV-2 in vitro. Cell Dis. 6:50. doi: 10.1038/s41421-020-00192-8Li, Z., Li, X., Huang, Y. Y., Wu, Y., Liu, R., Zhou, L., et al. (2020). Identify potent SARS-CoV-2 main protease inhibitors via accelerated free energy perturbation-based virtual screening of existing drugs. Proc. Natl. Acad. Sci. 117, 27381–27387. doi: 10.1073/pnas.2010470117Lotfi, M., Hamblin, M. R., and Rezaei, N. (2020). COVID-19: transmission, prevention, and potential therapeutic opportunities. Clin. Chim. Acta 508, 254–266. doi: 10.1016/j.cca.2020.05.044Lu, Y., Liu, D. X., and Tam, J. P. (2008). Lipid rafts are involved in SARS-CoV entry into Vero E6 cells. Biochem. Biophys. Res. Commun. 369, 344–349. doi: 10.1016/j.bbrc.2008.02.023Marín-Palma, D., Tabares-Guevara, J. H., Zapata-Cardona, M. I., Flórez-Álvarez, L., Yepes, L. M., Rugeles, M. T., et al. (2021). Curcumin inhibits in vitro SARS-CoV-2 infection in vero E6 cells through multiple antiviral mechanisms. Molecules 26:6900. doi: 10.3390/molecules26226900Martínez-Gutierrez, M., Castellanos, J. E., and Gallego-Gómez, J. C. (2011). Statins reduce dengue virus production via decreased virion assembly. Intervirology 54, 202–216. doi: 10.1159/000321892Matsushita, K., Ding, N., Kou, M., Hu, X., Chen, M., Gao, Y., et al. (2020). The relationship of COVID-19 severity with cardiovascular disease and its traditional risk factors: a systematic review and meta-analysis. Glob. Heart 15, 64–64. doi: 10.5334/gh.814Mehrbod, P., Ideris, A., Omar, A. R., and Hair-Bejo, M. (2012). Evaluation of antiviral effect of atorvastatin on H1N1 infection in MDCK cells. Afr. J. Microbiol. Res. 6, 5715–5719. doi: 10.5897/AJMR12.1011Mendonça, L., Howe, A, Gilchrist, JB, Sun, D, Knight, ML, Zanetti-Domingues, LC, et al., (2020). SARS-CoV-2 assembly and egress pathway revealed by correlative multi-modal multi-scale Cryo-imaging. bioRxiv [Preprint].Mendoza, E. J., Manguiat, K., Wood, H., and Drebot, M. (2020). Two Detailed Plaque Assay Protocols for the Quantification of Infectious SARS-CoV-2. Curr. Protoc. Microbiol. 57:ecpmc105. doi: 10.1002/cpmc.105Minz, M. M., Bansal, M., and Kasliwal, R. R. (2020). Statins and SARS-CoV-2 disease: current concepts and possible benefits. Diabetes Metab. Syndr. 14, 2063–2067. doi: 10.1016/j.dsx.2020.10.021Mukherjee, S., Bhattacharyya, D., and Bhunia, A. (2020). Host-membrane interacting interface of the SARS coronavirus envelope protein: immense functional potential of C-terminal domain. Biophys. Chem. 266, 106452–106452. doi: 10.1016/j.bpc.2020.106452Mycroft-West, C. J., Su, D., Pagani, I., Rudd, T. R., Elli, S., Gandhi, N. S., et al. (2020). Heparin inhibits cellular invasion by SARS-CoV-2: structural dependence of the interaction of the spike S1 receptor-binding domain with heparin. Thromb. Haemost. 120, 1700–1715. doi: 10.1055/s-0040- 1721319Naik, V. R., Munikumar, M., Ramakrishna, U., Srujana, M., Goudar, G., Naresh, P., et al. (2020). Remdesivir (GS-5734) as a therapeutic option of 2019-nCOV main protease - in silico approach. J. Biomol. Struct. Dyn. 39, 4701–4714. doi: 10.1080/07391102.2020.1781694Nimgampalle, M., Devanathan, V., and Saxena, A. (2020). Screening of Chloroquine, Hydroxychloroquine and its derivatives for their binding affinity to multiple SARS-CoV-2 protein drug targets. J. Biomol. Struct. Dyn. 39, 4949–4961. doi: 10.1080/07391102.2020.1782265Otto, S. P., Day, T., Arino, J., Colijn, C., Dushoff, J., Li, M., et al. (2021). The origins and potential future of SARS-CoV-2 variants of concern in the evolving COVID-19 pandemic. Curr. Biol. 31, R918–R929. doi: 10.1016/j. cub.2021.06.049https://doi.org/10.3389/fmolb.2020.578964Pan, H., Peto, R., Henao-Restrepo, A. M., Preziosi, M. P., Sathiyamoorthy, V., Abdool Karim, Q., et al. (2021). Repurposed antiviral drugs for Covid-19 - interim WHO solidarity trial results. N. Engl. J. Med. 384, 497–511. doi: 10.1056/NEJMoa2023184Parquet, V., Henry, M., Wurtz, N., Dormoi, J., Briolant, S., Gil, M., et al. (2010). Atorvastatin as a potential anti-malarial drug: in vitro synergy in combinational therapy with quinine against plasmodium falciparum. Malar. J. 9:139. doi: 10.1186/1475-2875-9-139Pawlos, A., Niedzielski, M., Gorzelak-Pabiś, P., Broncel, M., and Woźniak, E. (2021). COVID-19: direct and indirect mechanisms of statins. Int. J. Mol. Sci. 22, 4177. doi: 10.3390/ijms22084177PFIZER (2010). Pfizer announces European Union approval of a new form of lipitor (atorvastatin) for use in children. Available at: https://www.pfizer. com/news/press-release/press-release-detail/pfizer_announces_european_ union_approval_of_a_new_form_of_lipitor_atorvastatin_for_use_in_children (Accessed January, 2021).Reiner, Ž., Hatamipour, M., Banach, M., Pirro, M., al-Rasadi, K., Jamialahmadi, T., et al. (2020). Statins and the COVID-19 main protease: in silico evidence on direct interaction. Arch. Med. Sci. 16, 490–496. doi: 10.5114/aoms.2020.94655Rudrapal, M., Khairnar, S., and Jadhav, A. (2020). Drug Repurposing (DR): An Emerging Approach in Drug Discovery. London: IntechOpen. Sabatino, J., de Rosa, S., di Salvo, G., and Indolfi, C. (2020). Impact of cardiovascular risk profile on COVID-19 outcome. A meta-analysis. PLoS One 15:e0237131. doi: 10.1371/journal.pone.0237131Sacco, M. D., Ma, C., Lagarias, P., Gao, A., Townsend, J. A., Meng, X., et al. (2020). Structure and inhibition of the SARS-CoV-2 main protease reveal strategy for developing dual inhibitors against Mspro and cathepsin L. Sci. Adv. 6:eabe0751. doi: 10.1126/sciadv.abe0751Sanders, D. W., et al., (2020). SARS-CoV-2 requires cholesterol for viral entry and pathological syncytia formation. bioRxiv [preprint].Schmidt, N. M., Wing, P. A. C., McKeating, J. A., and Maini, M. K. (2020). Cholesterol-modifying drugs in COVID-19. Oxf. open immunol. 1:iqaa001. doi: 10.1093/oxfimm/iqaa001Schöning-Stierand, K., Diedrich, K., Fährrolfes, R., Flachsenberg, F., Meyder, A., Nittinger, E., et al. (2020). ProteinsPlus: interactive analysis of protein–ligand binding interfaces. Nucleic Acids Res. 48, W48–W53. doi: 10.1093/nar/gkaa235Shrivastava-Ranjan, P., Flint, M., Bergeron, É., McElroy, A. K., Chatterjee, P., Albariño, C. G., et al. (2018). Statins Suppress Ebola Virus Infectivity by Interfering with Glycoprotein Processing. MBio 9, e00660–e00618. doi: 10.1128/mBio.00660-18Simabuco, F. M., Tamura, R. E., Pavan, I. C. B., Morale, M. G., and Ventura, A. M. (2021). Molecular mechanisms and pharmacological interventions in the replication cycle of human coronaviruses. Genet. Mol. Biol. 44:e20200212. doi: 10.1590/1678-4685-gmb-2020-0212Singh, D. D., Han, I., Choi, E. H., and Yadav, D. K. (2020). Recent advances in pathophysiology, drug development and future perspectives of SARSCoV-2. Front. Cell Dev. Biol. 8:580202. doi: 10.3389/fcell.2020.580202Su, H. X., Yao, S., Zhao, W. F., Li, M. J., Liu, J., Shang, W. J., et al. (2020). Anti-SARS-CoV-2 activities in vitro of Shuanghuanglian preparations and bioactive ingredients. Acta Pharmacol. Sin. 41, 1167–1177. doi: 10.1038/ s41401-020-0483-6Su, H. X., Yao, S., Zhao, W. F., Li, M. J., Liu, J., Shang, W. J., et al. (2020). Anti-SARS-CoV-2 activities in vitro of Shuanghuanglian preparations and bioactive ingredients. Acta Pharmacol. Sin. 41, 1167–1177. doi: 10.1038/ s41401-020-0483-6Tandon, R., Sharp, J. S., Zhang, F., Pomin, V. H., Ashpole, N. M., Mitra, D., et al. (2021). Effective inhibition of SARS-CoV-2 entry by heparin and enoxaparin derivatives. J. Virol. 95, e01987–e01920. doi: 10.1128/ JVI.01987-20Trott, O., and Olson, A. J. (2010). AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 31, 455–461. doi: 10.1002/jcc.21334Uzunova, K., Filipova, E., Pavlova, V., and Vekov, T. (2020). Insights into antiviral mechanisms of remdesivir, lopinavir/ritonavir and chloroquine/ hydroxychloroquine affecting the new SARS-CoV-2. Biomed. Pharmacother. 131:110668. doi: 10.1016/j.biopha.2020.110668TUzunova, K., Filipova, E., Pavlova, V., and Vekov, T. (2020). Insights into antiviral mechanisms of remdesivir, lopinavir/ritonavir and chloroquine/ hydroxychloroquine affecting the new SARS-CoV-2. Biomed. Pharmacother. 131:110668. doi: 10.1016/j.biopha.2020.110668Villareal, V. A., Rodgers, M. A., Costello, D. A., and Yang, P. L. (2015). Targeting host lipid synthesis and metabolism to inhibit dengue and hepatitis C viruses. Antivir. Res. 124, 110–121. doi: 10.1016/j.antiviral.2015.10.013V’kovski, P., Kratzel, A., Steiner, S., Stalder, H., and Thiel, V. (2021). Coronavirus biology and replication: implications for SARS-CoV-2. Biomed. Pharmacother. 19, 155–121. doi: 10.1038/s41579-020-00468-6 Wang, M., Cao, R., Zhang, L., Yang, X., Liu, J., XWang, H., Yuan, Z, Pavel, M. A., Jablonski, S. M., Jablonski, J., Hobson, R., et al., (2021). The role of high cholesterol in age-related COVID19 lethality. bioRxiv [Preprint]. doi: 10.1101/2020.05.09.086249Wang, M., Cao, R., Zhang, L., Yang, X., Liu, J., Xu, M., et al. (2020). Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro. Cell Res. 30, 269–271. doi: 10.1038/ s41422-020-0282-0Wang, Y., Zhang, D., du, G., du, R., Zhao, J., Jin, Y., et al. (2020). Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial. Lancet 395, 1569–1578. doi: 10.1016/S0140-6736(20) 31022-9WHO (2020a). COVID-19: cronología de la actuación de la OMS. Available at: https://www.who.int/es/news/item/27-04-2020-who-timeline---covid-19 (Accessed January, 2022).WHO (2020b). WHO Coronavirus Disease (COVID-19) Dashboard. Available at: https://covid19.who.int/ (Accessed January, 2021).Wolff, G., Melia, C. E., Snijder, E. J., and Bárcena, M. (2020). Double-membrane vesicles as platforms for viral replication. Trends Microbiol. 28, 1022–1033. doi: 10.1016/j.tim.2020.05.009Wösten-van Asperen, R. M., Bos, A. P., Bem, R. A., Dierdorp, B. S., Dekker, T., van Goor, H., et al. (2013). Imbalance between pulmonary angiotensinconverting enzyme and angiotensin-converting enzyme 2 activity in acute respiratory distress syndrome. Pediatr. Crit. Care Med. 14, e438–e441. doi: 10.1097/PCC.0b013e3182a55735Wurtz, N., Penant, G., Jardot, P., Duclos, N., and la Scola, B. (2021). Culture of SARS-CoV-2 in a panel of laboratory cell lines, permissivity, and differences in growth profile. EJCDEU 40, 477–484. doi: 10.1007/ s10096-020-04106-0Yepes-Perez, A. F., Herrera-Calderón, O., Oliveros, C. A., Flórez-Álvarez, L., Zapata-Cardona, M. I., Yepes, L., et al. (2021). The Hydroalcoholic extractYuan, S., Chan, C. C. Y., Chik, K. K. H., Tsang, J. O. L., Liang, R., Cao, J., et al. (2020). Broad-Spectrum host-based antivirals targeting the interferon and lipogenesis pathways as potential treatment options for the pandemic coronavirus disease 2019 (COVID-19). Viruses 12:628. doi: 10.3390/ v12060628Zapatero-Belinchón, F. J., et al. (2021). Fluvastatin mitigates SARS-CoV-2 infection in human lung cells. iScience 24:103469. doi: 10.1016/j.isci.2021. 103469Zhang, Q., Chen, C. Z., Swaroop, M., Xu, M., Wang, L., Lee, J., et al. (2020). Heparan sulfate assists SARS-CoV-2 in cell entry and can be targeted by approved drugs in vitro. Cell Dis. 6:80. doi: 10.1038/s41421-020-00222-5PublicationORIGINALATV SARS 2022.pdfATV SARS 2022.pdfapplication/pdf1994444https://repository.ucc.edu.co/bitstreams/f7441486-babe-4131-890c-366a8f9e3998/download93c6a2a9cb98625ec4fa1b31c88155a0MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repository.ucc.edu.co/bitstreams/648d10dc-ed05-4306-a8f0-67d0f2b999a4/download8a4605be74aa9ea9d79846c1fba20a33MD52THUMBNAILATV SARS 2022.pdf.jpgATV SARS 2022.pdf.jpgGenerated Thumbnailimage/jpeg5844https://repository.ucc.edu.co/bitstreams/afbd3188-ac71-4bc4-b45c-fb5b51935df2/downloade171505baddf792665a539622779d725MD53TEXTATV SARS 2022.pdf.txtATV SARS 2022.pdf.txtExtracted texttext/plain88022https://repository.ucc.edu.co/bitstreams/0b94634a-975f-48e4-add7-12f7c942b97f/download59a33f910fb8fc94513c2e4ff101532bMD5420.500.12494/45190oai:repository.ucc.edu.co:20.500.12494/451902024-08-20 16:23:43.575open.accesshttps://repository.ucc.edu.coRepositorio Institucional Universidad Cooperativa de Colombiabdigital@metabiblioteca.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 |