Virucidal and antiviral activities of pomegranate (Punica granatum) extract against the mosquito-borne Mayaro virus

Tiago Souza Salles,Marcelo Damião Ferreira Meneses,Davis Fernandes Ferreira,Danielle M De Oliveira,Márcia Regina Soares,José A Ventura,Lucio Ayres Caldas,Ricardo M Kuster,Renata Campos Azevedo,Thayane Encarnação Sá-Guimarães

doi:10.1186/s13071-021-04955-4

Abstract

BackgroundThe arthropod-borne Mayaro virus (MAYV) causes “Mayaro fever,” a disease of medical significance, primarily affecting individuals in permanent contact with forested areas in tropical South America. Recently, MAYV has attracted attention due to its likely urbanization. There are currently no licensed drugs against most mosquito-transmitted viruses. Punica granatum (pomegranate) fruits cultivated in Brazil have been subjected to phytochemical investigation for the identification and isolation of antiviral compounds. In the present study, we explored the antiviral activity of pomegranate extracts in Vero cells infected with Mayaro virus.MethodsThe ethanol extract and punicalagin of pomegranate were extracted solely from the shell and purified by chromatographic fractionation, and were chemically identified using spectroscopic techniques. The cytotoxicity of the purified compounds was measured by the dye uptake assay, while their antiviral activity was evaluated by a virus yield inhibition assay.ResultsPomegranate ethanol extract (CC50 = 588.9, IC50 = 12.3) and a fraction containing punicalagin as major compound (CC50 = 441.5, IC50 = 28.2) were shown to have antiviral activity (SI 49 and 16, respectively) against Mayaro virus, an alphavirus. Immunofluorescence analysis showed the virucidal effect of pomegranate extract, and transmission electron microscopy (TEM) revealed damage in viral particles treated with this extract.ConclusionsThe P. granatum extract is a promising source of antiviral compounds against the alphavirus MAYV and represents an excellent candidate for future studies with other enveloped RNA viruses.Graphical abstract

Highlights

The arthropod-borne Mayaro virus (MAYV) causes “Mayaro fever,” a disease of medical significance, primarily affecting individuals in permanent contact with forested areas in tropical South America
Those (23–25) with a large purple spot on the thin-layer chromatography (TLC) plate sprayed with a sodium nitrite/acetic acid 10% solution were pooled and analyzed by high-performance liquid chromatography with diode array detection (HPLC/ DAD), electrospray ionization/mass spectrometry (ESI/MS), and nuclear magnetic resonance (NMR)
Structure identification of P. granatum isolate The chromatographic fractionation of the EtOH extract fruits led to isolation of 34 fractions, which were analyzed by TLC

Summary

Introduction

The arthropod-borne Mayaro virus (MAYV) causes “Mayaro fever,” a disease of medical significance, primarily affecting individuals in permanent contact with forested areas in tropical South America. We explored the antiviral activity of pomegranate extracts in Vero cells infected with Mayaro virus. Mayaro fever is a dengue-like but usually non-fatal illness, occurring in tropical South America and endemic to the Amazon region [1] where outbreaks have been registered. The virus was first isolated in Trinidad and Tobago [2], and imported cases of human infection occur, but not frequently, outside the Amazon region [3], some cases have been reported in other regions [4, 5]. Its genome has approximately 11 kilo-base pairs (kbp) coding for two polyproteins that are cleaved into non-structural proteins (nsP1, nsP2, snP3, and nsP4) and structural proteins (C, E2, E3, 6k, E1) [2]

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