This article presented the inhibitory activity of methyl 3, 4-dihydroxyphenylacetate on the enterovirus 71 (EV71) infection.  The EV71 VP1 capsid protein expression levels were analyzed with Western blotting.  Results revealed that the compound is able to inhibit EV71 replication in rhabdomyosarcoma (RD) cells.  After being incubated with the compound at a concentration of 0.01 μg·μL⁻¹ for 48 h, the level of EV71 vp1 mRNA in RD cells decreased by (76.83 ± 2.47) %.  The cytotoxic activity of the compound was evaluated against RD cells by a MTT assay.  The results showed that the compound had low toxicity with a CC₅₀ of  0.072 6 μg·μL⁻¹.  These findings suggest that methyl 3, 4-dihydroxyphenylacetate is a novel compound for antiviral therapies against EV71, which merited further investigation.