Pemetrexed inhibits Kaposi's sarcoma-associated herpesvirus replication through blocking dTMP synthesis.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. In immunocompromised patients, KSHV infection is capable of causing severe and fatal diseases. Current antiviral treatments for KSHV infections consist mostly of nucleoside analogs, all of which target viral polymerases and are associated with adverse effects and drug resistance. By screening an FDA-approved drug library, we identified pemetrexed as a potent anti-KSHV agent, with an IC50 of 90 nM. Characterization of the antiviral properties of pemetrexed revealed that it interferes with the lytic replication of viral DNA, resulting in the reduction of infectious virions. The antiviral effect of pemetrexed depends on the dTMP synthesis pathway that requires the folate-dependent enzymes. Besides, pemetrexed shows a broad spectrum of anti-herpes virus activity. Thus, our findings suggest that pemetrexed inhibits the lytic replication of KSHV DNA by blocking dTMP synthesis. Pemetrexed has the potential to be utilized as an anti-KSHV agent.