Mayaro virus (MAYV) is an emerging arbovirus with increasing circulation across the Americas. In the present study, we evaluated the potential antiviral activity of the following natural com-pounds against MAYV and other arboviruses: Sanguinarine, (R)-Shikonin, Fisetin, Honokiol, Tanshinone IIA and alfa-Mangostin. Sanguinarine and Shikonin showed significant cytotoxicity, whereas Fisetin, Honokiol, Tanshinone IIA and alfa-Mangostin were well-tolerated in all the cell lines tested. Honokiol and alfa-Mangostin treatment protected Vero-E6 cells against MAYV-induced damage and resulted in a dose-dependent reduction in viral progeny yields for each of the MAYV strains and human cell lines assessed. Also, Honokiol and alfa-Mangostin disrupted MAYV infection at different stages of the virus life cycle. Moreover, these compounds de-creased Una, Chikungunya and Zika viral titers and downmodulated the expression of E1 and nsP1 viral proteins from MAYV, Una and Chikungunya. Finally, in Honokiol- and alfa-Mangostin-treated cells, we observed an upregulation in the expression of type I interferon and specific interferon-stimulated genes, including IFN-alfa, IFN-beta;, MxA, ISG15, OAS2, MDA-5, TNF-alfa and IL-1beta;, which may promote an antiviral cellular state. Our results indicate that Honokiol and alfa-Mangostin present potential broad-spectrum activity against different arboviruses through a possible modulation of the interferon pathway.