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Planta Med 2020; 86(18): 1363-1374
DOI: 10.1055/a-1232-5705
Biological and Pharmacological Activity
Original Papers

Punica granatum Leaf Ethanolic Extract and Ellagic Acid as Inhibitors of Zika Virus Infection

Stefano Acquadro‡
1   Department of Drug Science and Technology, University of Turin, Turin, Italy
,
Andrea Civra‡
2   Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy
,
Cecilia Cagliero
1   Department of Drug Science and Technology, University of Turin, Turin, Italy
,
Arianna Marengo
1   Department of Drug Science and Technology, University of Turin, Turin, Italy
,
Massimo Rittà
2   Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy
,
Rachele Francese
2   Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy
,
Cinzia Sanna
3   Department of Environmental and Life Sciences University of Cagliari, Cagliari, Italy
,
Cinzia Bertea
4   Department of Life Sciences and Systems Biology, University of Turin, Turin, Italy
,
Barbara Sgorbini 
1   Department of Drug Science and Technology, University of Turin, Turin, Italy
,
David Lembo
2   Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy
,
Manuela Donalisio
2   Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy
,
Patrizia Rubiolo 
1   Department of Drug Science and Technology, University of Turin, Turin, Italy
› Author AffiliationsSupported by: Università degli Studi di Torino Ricerca Locale (ex-60% 2018)
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Abstract

Zika virus, an arthropod-borne flavivirus, is an emerging healthcare threat worldwide. Zika virus is responsible for severe neurological effects, such as paralytic Guillain-Barrè syndrome, in adults, and also congenital malformations, especially microcephaly. No specific antiviral drugs and vaccines are currently available, and treatments are palliative, but medicinal plants show great potential as natural sources of anti-Zika phytochemicals. This study deals with the investigation of the composition, cytotoxicity, and anti-Zika activity of Punica granatum leaf ethanolic extract, fractions, and phytoconstituents. P. granatum leaves were collected from different areas in Italy and Greece in different seasons. Crude extracts were analyzed and fractionated, and the pure compounds were isolated. The phytochemical and biomolecular fingerprint of the pomegranate leaves was determined. The antiviral activities of the leaf extract, fractions, and compounds were investigated against the MR766 and HPF2013 Zika virus strains in vitro. Both the extract and its fractions were found to be active against Zika virus infection. Of the compounds isolated, ellagic acid showed particular anti-Zika activities, with EC50 values of 30.86 µM for MR766 and 46.23 µM for HPF2013. The mechanism of action was investigated using specific antiviral assays, and it was demonstrated that ellagic acid was primarily active as it prevented Zika virus infection and was able to significantly reduce Zika virus progeny production. Our data demonstrate the anti-Zika activity of pomegranate leaf extract and ellagic acid for the first time. These findings identify ellagic acid as a possible anti-Zika candidate compound that can be used for preventive and therapeutic interventions.

Key words

Zika virus - Punica granatum - Lythraceae - leaf ethanolic extract - phytochemical and biomolecular fingerprint - antiviral - ellagic acid

Supporting Information

    Anti-ZIKV activity of a reference pomegranate leaf ethanolic extract PGL8 (Fig. 1S); cell viability assays (Fig. 2S); LC chromatograms of the pomegranate leaf extract PGL8 (Fig. 3S); GC-MS chromatograms of the pomegranate leaf extract PGL8 (Fig. 4S); comparison of ITS sequences between the 11 P. granatum samples (Fig. 5S); comparison of psbA-trnH sequences between the 11 P. granatum samples (Fig. 6S); anti-MR766 activity of PGSum85, PGSum95, PGAut85, and PGAut95 (Fig. 7S); 1H 600 MHz NMR of ellagic acid isolated with LC-Prep and of the commercial standard reference (Fig. 8S); inactivation of MR766 particles by isolated ellagic acid (Fig. 9S); effect of isolated ellagic acid on virus binding to cells (Fig. 10S); location, coordinates, and code of pomegranate leaf samples (Table 1S); concentration of phenolic compounds and triterpenes in the different pomegranate leaf extracts and in the fractions (Table 2S); list of the sequences obtained from P. granatum samples deposited in GenBank (Table 3S); list of primers used in PCR and sequencing (Table 4S); quantification method, linearity range, R2, and calibration curve of the main compounds by UV and SRM methods (Table 5S); and target ion, qualifier ions, linearity range, R2, and calibration curve of triterpenoids quantified by GC-MS (Table 6S) are available as Supporting Information.

  • Supporting Information


Publication History

Received: 22 May 2020

Accepted after revision: 30 July 2020

Article published online:
16 September 2020

© 2020. Thieme. All rights reserved.

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