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Infections caused by these viruses may cause long-term neurological sequelae in humans. The continuous emergence of infections caused by viruses around the world, such as the chikungunya virus (CHIKV) (Alphavirus genus), the zika virus (ZIKV) and the yellow fever virus (YFV) (both of the Flavivirus genus), warrants the development of new strategies to combat them. Our study demonstrates the inhibitory potential of the water-soluble vitamin riboflavin against NS2B\/NS3pro of ZIKV and YFV and nsP2pro of CHIKV. Riboflavin presents a competitive inhibition mode with IC50 values in the medium \u00b5M range of 79.4 \u00b1 5.0 \u00b5M for ZIKV NS2B\/NS3pro and 45.7 \u00b1 2.9 \u03bcM for YFV NS2B\/NS3pro. Against CHIKV nsP2pro, the vitamin showed a very strong effect (93 \u00b1 5.7 nM). The determined dissociation constants (KD) are significantly below the threshold value of 30 \u00b5M. The ligand binding increases the thermal stability between 4 \u00b0C and 8 \u00b0C. Unexpectedly, riboflavin showed inhibiting activity against another viral protein; the molecule was also able to inhibit the viral entry of CHIKV. Molecular dynamics simulations indicated great stability of riboflavin in the protease active site, which validates the repurposing of riboflavin as a promising molecule in drug development against the viruses presented here.<\/jats:p>","DOI":"10.3390\/microorganisms10071331","type":"journal-article","created":{"date-parts":[[2022,7,1]],"date-time":"2022-07-01T00:21:50Z","timestamp":1656634910000},"page":"1331","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Riboflavin, a Potent Neuroprotective Vitamin: Focus on Flavivirus and Alphavirus Proteases"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8763-3884","authenticated-orcid":false,"given":"Raphael J.","family":"Eberle","sequence":"first","affiliation":[{"name":"Institute of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum J\u00fclich GmbH, 52428 J\u00fclich, Germany"},{"name":"Institut f\u00fcr Physikalische Biologie, Heinrich-Heine-Universit\u00e4t D\u00fcsseldorf, Universit\u00e4tsstra\u00dfe, 40225 D\u00fcsseldorf, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Danilo S.","family":"Olivier","sequence":"additional","affiliation":[{"name":"Center of Integrated Sciences, Campus Cimba, Federal University of Tocantins, Aragua\u00edna 77824-838, TO, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8101-6933","authenticated-orcid":false,"given":"Marcos S.","family":"Amaral","sequence":"additional","affiliation":[{"name":"Institute of Physics, Federal University of Mato Grosso do Sul, Campo Grande 79070-900, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolina C.","family":"Pacca","sequence":"additional","affiliation":[{"name":"Instituto Superior de Educa\u00e7\u00e3o Ceres, FACERES Medical School, S\u00e3o Jos\u00e9 do Rio Preto 15090-305, SP, Brazil"},{"name":"Laborat\u00f3rio de Pesquisas em Virologia, Departamento de Doen\u00e7as Dermatol\u00f3gicas, Infecciosas e Parasit\u00e1rias, Faculdade de Medicina de S\u00e3o Jos\u00e9 do Rio Preto-FAMERP, S\u00e3o Jos\u00e9 do Rio Preto 15090-000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1102-2419","authenticated-orcid":false,"given":"Mauricio L.","family":"Nogueira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisas em Virologia, Departamento de Doen\u00e7as Dermatol\u00f3gicas, Infecciosas e Parasit\u00e1rias, Faculdade de Medicina de S\u00e3o Jos\u00e9 do Rio Preto-FAMERP, S\u00e3o Jos\u00e9 do Rio Preto 15090-000, SP, Brazil"},{"name":"Department of Pathology, University of Texas Medical Branch, Galveston, TX 77550, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2460-1145","authenticated-orcid":false,"given":"Raghuvir K.","family":"Arni","sequence":"additional","affiliation":[{"name":"Multiuser Center for Biomolecular Innovation, Department of Physics, IBILCE, S\u00e3o Paulo State University, S\u00e3o Jose do Rio Preto 15054-000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0065-7366","authenticated-orcid":false,"given":"Dieter","family":"Willbold","sequence":"additional","affiliation":[{"name":"Institute of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum J\u00fclich GmbH, 52428 J\u00fclich, Germany"},{"name":"Institut f\u00fcr Physikalische Biologie, Heinrich-Heine-Universit\u00e4t D\u00fcsseldorf, Universit\u00e4tsstra\u00dfe, 40225 D\u00fcsseldorf, Germany"},{"name":"JuStruct: J\u00fclich Centre for Structural Biology, Forchungszentrum J\u00fclich, 52428 J\u00fclich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6518-6497","authenticated-orcid":false,"given":"Monika A.","family":"Coronado","sequence":"additional","affiliation":[{"name":"Institute of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum J\u00fclich GmbH, 52428 J\u00fclich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"12040","DOI":"10.1128\/JVI.00133-07","article-title":"Neuroinvasion of fluorescein-positive monocytes in acute simian immunodeficiency virus infection","volume":"81","author":"Clay","year":"2007","journal-title":"J. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"8676","DOI":"10.1523\/JNEUROSCI.1860-10.2010","article-title":"A novel population of myeloid cells responding to coxsackievirus infection assists in the dissemination of virus within the neonatal CNS","volume":"30","author":"Ruller","year":"2010","journal-title":"J. Neurosci."},{"key":"ref_3","unstructured":"Schmaljohn, A.L., McClain, D., and Baron, S. (1996). Chapter 54 Alphaviruses (Togaviridae) and Flaviviruses (Flaviviridae). Medical Microbiology, University of Texas Medical Branch at Galveston. [4th ed.]."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/S0065-3527(03)60009-6","article-title":"Pathogenesis and pathophysiology of yellow fever","volume":"60","author":"Monath","year":"2003","journal-title":"Adv. Virus Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"815","DOI":"10.4269\/ajtmh.1963.12.815","article-title":"Histopathology of the central nervous system in swiss mice intracerebrally inoculated with 17-D strain of yellow fever virus","volume":"12","author":"Dominguez","year":"1963","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1038\/s41541-022-00460-6","article-title":"Impact of yellow fever virus envelope protein on wild-type and vaccine epitopes and tissue tropism","volume":"7","author":"Davis","year":"2022","journal-title":"Npj Vaccines"},{"key":"ref_7","first-page":"145","article-title":"West Nile virus infection and its neurological manifestations","volume":"1","author":"Madden","year":"2003","journal-title":"J. Clin. Med. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2107","DOI":"10.1128\/JVI.75.5.2107-2118.2001","article-title":"Yellow fever virus encephalitis: Properties of the brain-associated T-cell response during virus clearance in normal and gamma interferon-deficient mice and requirement for CD4+ lymphocytes","volume":"75","author":"Liu","year":"2001","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5602","DOI":"10.1128\/JVI.00143-15","article-title":"Viral infection of the central nervous system and neuroinflammation precede blood-brain barrier disruption during Japanese encephalitis virus infection","volume":"89","author":"Li","year":"2015","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1186\/s12974-017-0837-2","article-title":"Development of a model of Saint Louis encephalitis infection and disease in mice","volume":"14","author":"Marques","year":"2017","journal-title":"J. Neuroinflammation"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"430","DOI":"10.12998\/wjcc.v3.i5.430","article-title":"Tick-borne encephalitis: A review of epidemiology, clinical characteristics, and management","volume":"3","author":"Bogovic","year":"2015","journal-title":"World J. Clin. Cases"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1007\/s00381-016-3074-6","article-title":"Microcephaly and Zika virus: Neonatal neuroradiological aspects","volume":"32","author":"Cavalheiro","year":"2016","journal-title":"Childs Nerv. Syst."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1531","DOI":"10.1016\/S0140-6736(16)00562-6","article-title":"Guillain-Barr\u00e9 Syndrome outbreak associated with Zika virus infection in French Polynesia: A case-control study","volume":"387","author":"Blake","year":"2016","journal-title":"Lancet"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e1978","DOI":"10.1002\/rmv.1978","article-title":"The neurological complications of chikungunya virus: A systematic review","volume":"28","author":"Mehta","year":"2018","journal-title":"Rev. Med. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"11490","DOI":"10.1128\/JVI.05379-11","article-title":"Alphavirus-induced encephalomyelitis: Antibody-secreting cells and viral clearance from the nervous system","volume":"85","author":"Metcalf","year":"2011","journal-title":"J. Virol."},{"key":"ref_16","first-page":"861912","article-title":"The Pathogenesis of Alphaviruses","volume":"2013","author":"Atkins","year":"2013","journal-title":"ISRN Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1352","DOI":"10.1093\/ajcn\/77.6.1352","article-title":"Riboflavin (vitamin B-2) and health","volume":"77","author":"Powers","year":"2003","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3159","DOI":"10.1128\/JCM.37.10.3159-3166.1999","article-title":"Molecular analysis of riboflavin synthesis genes in Bartonella henselae and use of the ribC gene for differentiation of Bartonella species by PCR","volume":"37","author":"Bereswill","year":"1999","journal-title":"J. Clin. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1097\/01.tp.0000256283.06469.d4","article-title":"Riboflavin-mediated reduction of oxidant injury, rejection, and vasculopathy after cardiac allotransplantation","volume":"83","author":"Iwanaga","year":"2007","journal-title":"Transplantation"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1820","DOI":"10.1128\/IAI.72.3.1820-1823.2004","article-title":"Highly purified riboflavin presents a promising therapeutic strategy for sepsis and septic shock","volume":"72","author":"Toyosawa","year":"2004","journal-title":"Infect. Immun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1571","DOI":"10.1126\/science.290.5496.1571","article-title":"Biochemical basis of oxidative protein folding in the endoplasmic reticulum","volume":"290","author":"Tu","year":"2000","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"30490","DOI":"10.1155\/MI\/2006\/30490","article-title":"Theaflavin ameliorates cerebral ischemia-reperfusion injury in rats through its anti-inflammatory effect and modulation of STAT-1","volume":"5","author":"Cai","year":"2006","journal-title":"Mediat. Inflamm."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1177\/1533210110392943","article-title":"Vitamin B2: Riboflavin","volume":"16","author":"Buehler","year":"2011","journal-title":"Evid. Based Complement. Alternat. Med."},{"key":"ref_24","unstructured":"Somer, E. (1995). The Essential Guide to Vitamins and Minerals, HarperCollins. [2nd ed.]."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1016\/S0140-6736(16)00320-2","article-title":"Zika virus and microcephaly: Why is this situation a PHEIC?","volume":"387","author":"Heymann","year":"2016","journal-title":"Lancet"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1038\/nm.4184","article-title":"Identification of small-molecule inhibitors of Zika virus infection and induced neural cell death via a drug repurposing screen","volume":"22","author":"Xu","year":"2016","journal-title":"Nat. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1126\/science.aag0219","article-title":"Countering the Zika epidemic in Latin America","volume":"353","author":"Ferguson","year":"2016","journal-title":"Science"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s12985-019-1277-7","article-title":"Recent sylvatic yellow fever virus transmission in Brazil: The news from an old disease","volume":"17","author":"Silva","year":"2020","journal-title":"Virol. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.3389\/fmicb.2020.01297","article-title":"Chikungunya Virus: An Emergent Arbovirus to the South American Continent and a Continuous Threat to the World","volume":"11","author":"Cunha","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.antiviral.2017.07.007","article-title":"Structure-based discovery of clinically approved drugs as Zika virus NS2B-NS3 protease inhibitors that potently inhibit Zika virus infection in vitro and in vivo","volume":"145","author":"Yuan","year":"2017","journal-title":"Antivir. Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Pathak, N., Lai, M.L., Chen, W.Y., Hsieh, B.W., Yu, G.Y., and Yang, J.M. (2017). Pharmacophore anchor models of flaviviral NS3 proteases lead to drug repurposing for DENV infection. BMC Bioinform., 18.","DOI":"10.1186\/s12859-017-1957-5"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2404","DOI":"10.1080\/22221751.2020.1838954","article-title":"Methylene blue is a potent and broad-spectrum inhibitor against Zika virus in vitro and in vivo","volume":"9","author":"Li","year":"2020","journal-title":"Emerg. Microbes Infect."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2616","DOI":"10.1021\/acsinfecdis.0c00217","article-title":"JMX0207, a Niclosamide Derivative with Improved Pharmacokinetics, Suppresses Zika Virus Infection Both In Vitro and In Vivo","volume":"6","author":"Li","year":"2020","journal-title":"ACS Infect. Dis."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/j.antiviral.2017.12.018","article-title":"Erythrosin B is a potent and broad-spectrum orthosteric inhibitor of the flavivirus NS2B-NS3 protease","volume":"150","author":"Li","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2223","DOI":"10.1099\/vir.0.82735-0","article-title":"Yellow fever virus NS3 protease: Peptide-inhibition studies","volume":"88","author":"Loehr","year":"2007","journal-title":"J. Gen. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Eberle, R.J., Olivier, D.S., Amaral, M.S., Willbold, D., Arni, R.K., and Coronado, M.A. (2021). Promising Natural Compounds against Flavivirus Proteases: Citrus Flavonoids Hesperetin and Hesperidin. Plants, 10.","DOI":"10.3390\/plants10102183"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Eberle, R.J., Olivier, D.S., Pacca, C.C., Avilla, C.M., Nogueira, M.L., Amaral, M.S., Willbold, D., Arni, R.K., and Coronado, M.A. (2021). In vitro study of Hesperetin and Hesperidin as inhibitors of zika and chikungunya virus proteases. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0246319"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5786","DOI":"10.1021\/acsomega.0c06191","article-title":"1,3-Thiazolbenzamide Derivatives as Chikungunya Virus nsP2 Protease Inhibitors","volume":"6","author":"Ivanova","year":"2021","journal-title":"ACS Omega"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"9791","DOI":"10.1021\/acsomega.1c00447","article-title":"Inhibition of Chikungunya Virus Infection by 4-Hydroxy-1-Methyl-3-(3-morpholinopropanoyl) quinoline-2 (1 H)-one (QVIR) Targeting nsP2 and E2 Proteins","volume":"6","author":"Islamuddin","year":"2021","journal-title":"ACS Omega"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1128\/mr.58.3.491-562.1994","article-title":"The alphaviruses: Gene expression, replication, and evolution","volume":"58","author":"Strauss","year":"1994","journal-title":"Microbiol. Rev."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1038\/nsmb1073","article-title":"Structural basis for the activation of flaviviral NS3 proteases from dengue and West Nile virus","volume":"13","author":"Erbel","year":"2006","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3135","DOI":"10.1007\/s11164-013-1159-3","article-title":"Analysis of binding interaction between vitamin B 2 and trypsin. Res","volume":"40","author":"Gao","year":"2014","journal-title":"Chem. Intermed."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"5847","DOI":"10.1021\/bi9529972","article-title":"Flavins inhibit human cytomegalovirus UL80 protease via disulfide bond formation","volume":"35","author":"Baum","year":"1996","journal-title":"Biochemistry"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.antiviral.2018.10.019","article-title":"Zika virus NS2B\/NS3 proteinase: A new target for an old drug-Suramin a lead compound for NS2B\/NS3 proteinase inhibition","volume":"160","author":"Coronado","year":"2018","journal-title":"Antiviral Res."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Roy, A., Lim, L., Srivastava, S., Lu, Y., and Song, J. (2017). Solution conformations of Zika NS2B-NS3pro and its inhibition by natural products from edible plants. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0180632"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Motulsky, H.J., and Christopoulos, A. (2003). Fitting Models to Biological Data Using Linear and Nonlinear Regression: A Practical Guide to Curve Fitting, GraphPad Software Inc.","DOI":"10.1093\/oso\/9780195171792.001.0001"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1016\/S1359-6446(04)03069-7","article-title":"Ligand efficiency: A useful metric for lead selection","volume":"9","author":"Hopkins","year":"2004","journal-title":"Drug Discov. Today"},{"key":"ref_48","first-page":"1104","article-title":"Tool development for Prediction of pIC50 values from the IC50 values\u2014A pIC50 value calculator","volume":"5","author":"Selvaraj","year":"2011","journal-title":"Curr. Trends Biotechnol. Pharm."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/S0076-6879(85)17018-7","article-title":"Nonlinear least-squares analysis","volume":"117","author":"Johnson","year":"1985","journal-title":"Meth. Enzymol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.dyepig.2005.11.008","article-title":"A study of the interaction between bromopyrogallol red and bovine serum albumin by spectroscopic methods","volume":"73","author":"Shaikh","year":"2007","journal-title":"Dye. Pigment."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.colsurfb.2015.07.063","article-title":"Probing the binding of trypsin to glutathione-stabilized gold nanoparticles in aqueous solution","volume":"135","author":"Wang","year":"2015","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2067","DOI":"10.1039\/C7AN00288B","article-title":"Association models for binding of molecules to nanostructures","volume":"142","author":"Ahumada","year":"2017","journal-title":"Analyst"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1016\/j.ijbiomac.2018.05.007","article-title":"Crystal structure of chikungunya virus nsP2 cysteine protease reveals a putative flexible loop blocking its active site","volume":"116","author":"Narwal","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.tmaid.2016.05.004","article-title":"The arboviral burden of disease caused by co-circulation and co-infection of Dengue, Chikungunya and Zika in the Americas","volume":"14","year":"2016","journal-title":"Travel Med. Infect. Dis."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1126\/science.aag2419","article-title":"Crystal structure of Zika virus NS2B-NS3 protease in complex with a boronate inhibitor","volume":"353","author":"Lei","year":"2016","journal-title":"Science"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading","volume":"31","author":"Trott","year":"2009","journal-title":"J. Comput. Chem."},{"key":"ref_57","unstructured":"Frisch, M.J., Trucks, G.W., Schlegel, H.B., Scuseria, G.E., Robb, M.A., Cheeseman, J.R., Scalmani, G., Barone, V., Mennucci, B., and Petersson, G.A. (2022, April 25). Gaussian 16, Available online: https:\/\/gaussian.com\/."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.jmgm.2005.12.005","article-title":"Automatic atom type and bond type perception in molecular mechanical calculations","volume":"25","author":"Wang","year":"2006","journal-title":"J. Mol. Graph. Model."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1002\/jcc.20035","article-title":"Development and testing of a general amber force field","volume":"25","author":"Wang","year":"2004","journal-title":"J. Comput. Chem."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1668","DOI":"10.1002\/jcc.20290","article-title":"The Amber biomolecular simulation programs","volume":"26","author":"Case","year":"2005","journal-title":"J. Comput. Chem."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1021\/acs.jctc.9b00591","article-title":"ff19SB: Amino-Acid-Specific Protein Backbone Parameters Trained against Quantum Mechanics Energy Surfaces in Solution","volume":"16","author":"Tian","year":"2020","journal-title":"J. Chem. Theory Comput."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"W368","DOI":"10.1093\/nar\/gki464","article-title":"H++: A server for estimating pKas and adding missing hydrogens to macromolecules","volume":"33","author":"Gordon","year":"2005","journal-title":"Nucleic Acids Res."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"3084","DOI":"10.1021\/ct400341p","article-title":"III. PTRAJ and CPPTRAJ: Software for processing and analysis of molecular synamics trajectory data","volume":"9","author":"Roe","year":"2013","journal-title":"J. Chem. Theory Com."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"2020","DOI":"10.1021\/ct3010485","article-title":"Improved generalized born solvent model parameters for protein simulations","volume":"9","author":"Nguyen","year":"2013","journal-title":"J. Chem. Theory Comput."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1126\/science.aaf5036","article-title":"Zika virus in the Americas: Early epidemiological and genetic findings","volume":"352","author":"Faria","year":"2016","journal-title":"Science"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"906","DOI":"10.3201\/eid2105.141727","article-title":"East\/Central\/South African genotype chikungunya virus, Brazil, 2014","volume":"21","author":"Teixeira","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Bulerson, F.G., Chambers, T.M., and Wiedbrauk, D.L. (1992). Plaque Assays. Virology: A Laboratory Manual, Academic Press, Inc.","DOI":"10.1016\/B978-0-12-144730-4.50019-9"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1097\/MOG.0b013e32834172c0","article-title":"Vitamins as drugs: The importance of pharmacokinetics in oral dosing","volume":"27","author":"Alpers","year":"2011","journal-title":"Curr. Opin. Gastroenterol."},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Lykkesfeldt, J., and Tveden-Nyborg, P. (2019). The pharmacokinetics of vitamin C. Nutrients, 11.","DOI":"10.3390\/nu11102412"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.biochi.2015.11.006","article-title":"Absorption and blood\/cellular transport of folate and cobalamin: Pharmacokinetic and physiological considerations","volume":"126","author":"Alpers","year":"2016","journal-title":"Biochimie"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"132652","DOI":"10.1016\/j.molstruc.2022.132652","article-title":"In silico detection of potential inhibitors from vitamins and their derivatives compounds against SARS-CoV-2 main protease by using molecular docking, molecular dynamic simulation and ADMET profiling","volume":"1258","author":"Belhassan","year":"2022","journal-title":"J. Mol. Struct."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1002\/bmb.2002.494030030035","article-title":"Protein tryptophan accessibility studied by fluorescence quenching","volume":"30","author":"Denicola","year":"2002","journal-title":"Biochem. Mol. Biol. Educ."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"13410","DOI":"10.1038\/ncomms13410","article-title":"Structure of the NS2B-NS3 protease from Zika virus after self-cleavage","volume":"7","author":"Phoo","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/0166-3542(90)90015-Y","article-title":"Virucidal activity of hypericin against enveloped and non-enveloped DNA and RNA viruses","volume":"13","author":"Tang","year":"1990","journal-title":"Antivir. Res."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1007\/s00705-019-04172-6","article-title":"Expression, purification and virucidal activity of two recombinant isoforms of phospholipase A2 from Crotalus durissus terrificus venom","volume":"164","author":"Russo","year":"2019","journal-title":"Arch. Virol."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1078\/094471103322331467","article-title":"Virucidal effect of peppermint oil on the enveloped viruses herpes simplex virus type 1 and type 2 in vitro","volume":"10","author":"Schuhmacher","year":"2003","journal-title":"Phytomedicine"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1038\/s41467-018-08138-1","article-title":"Virucidal nano-perforator of viral membrane trapping viral RNAs in the endosome","volume":"10","author":"Kong","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1016\/j.bpj.2017.06.061","article-title":"Effects of HIV-1 gp41-Derived Virucidal Peptides on Virus-like Lipid Membranes","volume":"113","author":"Carravilla","year":"2017","journal-title":"Biophys. J."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"129521","DOI":"10.1016\/j.bbagen.2020.129521","article-title":"Structural characterization and polymorphism analysis of the NS2B-NS3 protease from the 2017 Brazilian circulating strain of Yellow Fever virus","volume":"1864","author":"Noske","year":"2020","journal-title":"Biochim. Biophys. Acta Gen. Subj."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"2348","DOI":"10.1128\/JCM.00877-16","article-title":"Coinfections of Zika and Chikungunya viruses in Bahia, Brazil, identified by metagenomic next-generation sequencing","volume":"54","author":"Sardi","year":"2016","journal-title":"J. Clin. Microbiol."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.ijid.2016.07.017","article-title":"Zika, dengue, and chikungunya co-infection in a pregnant woman from Colombia","volume":"51","author":"Castellanos","year":"2016","journal-title":"Int. J. Infect. Dis."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"42","DOI":"10.3389\/fmicb.2017.00042","article-title":"Zika virus, chikungunya virus, and dengue virus in cerebrospinal fluid from adults with neurological manifestations, Guayaquil, Ecuador","volume":"8","author":"Acevedo","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_83","doi-asserted-by":"crossref","unstructured":"Carrillo-Hern\u00e1ndez, M.Y., Ruiz-Saenz, J., Villamizar, L.J., G\u00f3mez-Rangel, S.Y., and Mart\u00ednez-Gutierrez, M. (2018). Co-circulation and simultaneous co-infection of dengue, chikungunya, and zika viruses in patients with febrile syndrome at the Colombian-Venezuelan border. BMC Infect. Dis., 18.","DOI":"10.1186\/s12879-018-2976-1"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/j.jiph.2018.09.007","article-title":"Co-infection between Zika and different Dengue serotypes during DENV outbreak in Brazil","volume":"12","author":"Estofolete","year":"2019","journal-title":"J. Infect. Public Health."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1099\/acmi.0.000300","article-title":"Dengue 2 serotype and yellow fever coinfection","volume":"3","author":"Gehrke","year":"2021","journal-title":"Access Microbiol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/7\/1331\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:41:33Z","timestamp":1760139693000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/10\/7\/1331"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,30]]},"references-count":85,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["microorganisms10071331"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms10071331","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,30]]}}}