A common childhood affliction of viral origin in young children and immunocompromised adults, the Hand, Foot and Mouth Disease (HFMD) has become a significant public health concern in the Asia-Pacific Region. Characterized by the appearance of vesiculopapular rashes on the hands, feet and mouth, the disease is generally mild and self-limiting. In a minority of cases, patients can develop neurological complications that could result in permanent morbidity or even fatality. In the absence of a specific antiviral for treatment, medical care is limited to supportive and symptomatic relief, presenting a need for more research into an effective antiviral to be used in the management of the disease. In this study, we evaluated the efficacy of chloroquine, a FDA-approved lysosomotropic agent, against several serotypes of HFMD-associated enteroviruses, including EV-A71, in reducing infectious virus production. We have also evaluated chloroquine in a murine model of EV-A71 infection to ascertain its antiviral efficacy in vivo. The results suggest that chloroquine could be a broad-acting antiviral effective against HFMD-associated enteroviruses.