Arboviruses, especially Dengue virus (DENV) and Zika virus (ZIKV), have been a severe threat to human health in the last few years due to uncontrollable transmission. There are no approved vaccines or clinical drugs available for use to prevent and treat their infections. Transient receptor potential mucolipin 2 and 3 (TRPML2 and TRPML3) were reported to modulate viral entry, but the antiviral function of these modulators was unknown. Here, we reported that ML-SA1, a TRPML agonist, inhibited DENV2 and ZIKV in vitro in a dose-dependent manner. Time-of-drug-addition experiments showed that ML-SA1 mainly restricted viral entry. Moreover, the selective TRPML3 activator SN-2 was found to share a similar antiviral effect against DENV2 and ZIKV, but the specific TRPML1 agonist MK6-83 was not effective. Although ML-SA1 was further revealed to induce autophagy, its antiviral role was independent of autophagy induction. Finally, ML-SA1 was found to inhibit DENV2 and ZIKV by promoting lysosome acidification and protease activity to cause viral degradation. Together, our study identifies two TRPML agonists, ML-SA1 and SN-2, as potent inhibitors of DENV2 and ZIKV, which may lead to the discovery of new candidates against viruses.

虫媒病毒，尤其是登革热病毒（DENV）和寨卡病毒（ZIKV），由于无法控制的传播，在过去几年中已严重威胁人类健康。没有批准的疫苗或临床药物可用于预防和治疗其感染。据报道，瞬时受体潜在的粘蛋白2和3（TRPML2和TRPML3）可以调节病毒的进入，但是这些调节剂的抗病毒功能尚不清楚。在这里，我们报道了TRPML激动剂ML-SA1在体外抑制DENV2和ZIKV以剂量依赖的方式。加药时间实验表明ML-SA1主要限制病毒进入。此外，发现选择性TRPML3激活剂SN-2对DENV2和ZIKV具有相似的抗病毒作用，但特异性TRPML1激动剂MK6-83无效。尽管进一步揭示了ML-SA1诱导自噬，但其抗病毒作用与自噬诱导无关。最后，发现ML-SA1通过促进溶酶体酸化和蛋白酶活性引起病毒降解来抑制DENV2和ZIKV。总之，我们的研究确定了两种TRPML激动剂ML-SA1和SN-2作为DENV2和ZIKV的有效抑制剂，这可能导致发现新的抗病毒候选物。