Viral methyltransferase inhibitors: berbamine, venetoclax, and ponatinib as efficacious antivirals against Chikungunya virus

Abstract

AbstractChikungunya virus (CHIKV), transmitted by mosquitoes, poses a significant global health threat. Presently, no effective treatment options are available to reduce the disease burden. The lack of approved therapeutics against CHIKV and the complex spectrum of chronic musculoskeletal and neurological manifestations raise significant concerns, and repurposing drugs could offer swift avenues in the development of effective treatment strategies. RNA capping is a crucial step meditated by non-structural protein 1 (nsP1) in CHIKV replication. In this study, FDA-approved antivirals targeting CHIKV nsP1 methyltransferase (MTase) have been identified by structure-based virtual screening. Berbamine Hydrochloride (BH), ABT199/Venetoclax (ABT), and Ponatinib (PT) were the top hits, which exhibited robust binding energies. Tryptophan fluorescence spectroscopy-based assay confirmed binding of BH-, ABT-, and PT to purified nsP1 with KDvalues ∼5.45 μM, ∼161.3 μM, and ∼3.83μM, respectively. A dose-dependent decrease in CHIKV nsP1 MTase activity was observed in a capillary electrophoresis-based assay. Treatment with BH, ABT, and PT lead to a dose-dependent reduction in the virus titer with IC50<100, ∼3.46, and <3.9 nM, respectively, and reduced viral mRNA levels. The nsP1 MTases are highly conserved among alphaviruses; therefore, BH, ABT, and PT, as expected, inhibited replication machinery in Sindbis virus (SINV) replicon assay with IC50∼1.94, ∼0.23, and >1.25 μM, respectively. These results underscore the efficacy and promise of repurposing drugs as rapid and effective antiviral therapeutics against CHIKV.HighlightsBerbamine Hydrochloride (BH), ABT199/Venetoclax (ABT), and Ponatinib (PT) identified as inhibitors of Chikungunya virus (CHIKV) nsP1Molecules BH, ABT and PT make moelcular interactions with nsP1: KDvalues ∼5.45, ∼161.3, and ∼3.83 in μM, respectively and effectively inhibit methyltransferase activity.BH, ABT, and PT exhibited potent antiviral activity against CHIKV in cell-based assays, with IC50<100, ∼3.46, and <3.9 nM.BH, ABT, and PT also inhibit Sindbis virus, the prototype alphavirus with IC50∼1.94, ∼0.23, and >1.25 μM respectively demonstrating broad spectrum antiviral efficacy.Graphical Abstract