<p>Background: Influenza virus, associated with high level of morbidity and mortality, has been recently considered a public health concern while the choices for the control and treatment of the disease are limited. The present study was conducted to evaluate activity of pomegranate peel extract and its fractions against <em>Influenza A</em> virus <em>in vitro</em>.&nbsp;<br /> Methods: In this research, ethyl alcohol extract of pomegranate peel was prepared and subjected to fractionation with different polarities. The potential <em>in vitro</em> anti-influenza A virus activity of the extract and fractions was assessed using Cytopathic Effect (CPE) reduction assay, Hemagglutinin Assay (HA), and 50% Tissue Culture In-fectious Doses (TCID₅₀) method in Madin-Darby Canine Kidney (MDCK) cells.<br /> Results: The crude pomegranate peel extract and its n-butanol and ethyl acetate fractions had the highest inhibitory effect against influenza A virus with IC₅₀ value of 6.45, 6.07 and 5.6 <em>&mu;g/ml</em> in MDCK cells, respectively. Our results also showed that, the production of virus was significantly reduced upon treatment with crude extract, n-butanol and ethyl acetate fractions in a dose-dependent manner (p&lt;0.05).<br /> Conclusion: Based on our results, the ethyl alcohol extract and its polar fractions of pomegranate peel can inhibit influenza A virus replication<em> in vitro</em>. Therefore, further characterization of its active ingredients and the mechanism of action should be carried out.&nbsp;</p>