Influenza A virus (IAV) infection is one of the major global public health concerns due to the continual emergence of new IAV strains with pandemic potential. Antivirals can be used as an intervention to combat IAV infection, however the increasing resistance to IAV-specific antivirals (e.g. Neuraminidase inhibitors and M2 ion channel inhibitors) demonstrates the need to develop novel antiviral therapies, with potentially different mechanisms of action. Polyphenol rich sugarcane extract (PRSE) is an extract prepared from the molasses of a sugarcane plant and in this study, we evaluated the antiviral activity of PRSE against different strains of IAV in vitro. We showed that PRSE treatment of IAV-infected MDCK cells inhibited IAV infectious virus production in a dose-dependent manner for multiple strains of the H3N2 and H1N1 subtype. Western blot and flow cytometry analysis revealed that PRSE treatment of IAV-infected MDCK cells at 2h post infection reduced viral protein expression. In addition, qRT-PCR showed that PRSE treatment modulated IAV viral genome transcription during infection. Haemagglutination inhibition assay and electron microscopy showed that PRSE treatment of infectious virions did not attenuate the haemagglutination ability or alter the morphology of virions. Using time of addition experiments, we observed that PRSE demonstrated antiviral activity against the early stages of IAV replication in vitro. Our recent data also indicates that the antiviral activity of PRSE is likely to be mediated through the attenuation of cell entry. In conclusion, PRSE is a potential antiviral candidate for IAV infection, and the antiviral activity against other viruses is to be investigated.