Abstract
Mayaro fever, caused by Mayaro virus (MAYV) is a sub-lethal disease with symptoms that are easily confused with those of dengue fever, except for polyarthralgia, which may culminate in physical incapacitation. Recently, outbreaks of MAYV have been documented in metropolitan areas, and to date, there is no therapy or vaccine available. Moreover, there is no information regarding the three-dimensional structure of the viral proteins of MAYV, which is important in the search for antivirals. In this work, we constructed a three-dimensional model of protein C of MAYV by homology modelling, and this was employed in a manner similar to that of receptors in virtual screening studies to evaluate 590 molecules as prospective antiviral agents. In vitro bioassays were utilized to confirm the potential antiviral activity of the flavonoid epicatechin isolated from Salacia crassifolia (Celastraceae). The virtual screening showed that six flavonoids were promising ligands for protein C. The bioassays showed potent antiviral action of epicatechin, which protected the cells from almost all of the effects of viral infection. An effective concentration (EC50) of 0.247 μmol/mL was observed with a selectivity index (SI) of 7. The cytotoxicity assay showed that epicatechin has low toxicity, with a 50% cytotoxic concentration (CC50) greater than 1.723 µmol/mL. Epicatechin was found to be twice as potent as the reference antiviral ribavirin. Furthermore, a replication kinetics assay showed a strong inhibitory effect of epicatechin on MAYV growth, with a reduction of at least four logs in virus production. Our results indicate that epicatechin is a promising candidate for further testing as an antiviral agent against Mayaro virus and other alphaviruses.
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Acknowledgements
We thank Brazilian Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the granting of Scientific Initiation grants. We thank the Universidade Federal de São João del-Rei for granting Master’s degree grants and the facilities to carry out this work.
Funding
This work was funded by the Brazilian Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) CBBAPQ01028-14.
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Supplementary material 2 Binding energy of the flavonoids and dioxanes docked to the protein C of the MAYV. *Molecules used as parameters for the evaluation of the binding energies obtained in Virtual Screening study with the Protein C of the MAYV (PDF 82 kb)
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Supplementary material 3 NMR spectrum of epicatechin from the extract of ethyl acetate from leaves of Salacia crassifolia. (a) 1H NMR (CD3OD, 400 MHz). (b) 13C NMR (CD3OD, 100 MHz) and the chemical structure of epicatechin (PDF 224 kb)
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Ferreira, P.G., Ferraz, A.C., Figueiredo, J.E. et al. Detection of the antiviral activity of epicatechin isolated from Salacia crassifolia (Celastraceae) against Mayaro virus based on protein C homology modelling and virtual screening. Arch Virol 163, 1567–1576 (2018). https://doi.org/10.1007/s00705-018-3774-1
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DOI: https://doi.org/10.1007/s00705-018-3774-1