基孔肯雅病毒(CHIKV),通过蚊子传播,对全球健康构成重大威胁。目前,目前尚无有效的治疗方案来减轻疾病负担.缺乏针对CHIKV的批准疗法以及复杂的慢性肌肉骨骼和神经系统表现引起了人们的关注。重新利用药物可以为制定有效的治疗策略提供迅速的途径。RNA加帽是非结构蛋白1(nsP1)在CHIKV复制中的关键步骤。在这项研究中,FDA批准的针对CHIKVnsP1甲基转移酶(MTase)的抗病毒药物已通过基于结构的虚拟筛选进行鉴定。盐酸小檗碱(BH),ABT199/维奈托克(ABT),和波纳替尼(PT)是最热门的,表现出强大的结合能。基于色氨酸荧光光谱法的测定证实了BH-,ABT-,和PT纯化nsP1,KD值为5.45μM,161.3μM,和〜3.83μM,分别。在基于毛细管电泳的检测中,观察到CHIKVnsP1MTase活性呈剂量依赖性下降.用BH治疗,ABT,和PT导致病毒滴度的剂量依赖性降低,IC50<100,〜6.75和<3.9nM,分别,并降低病毒mRNA水平。nsP1MTases在甲病毒中高度保守;因此,BH,ABT,还有PT,正如预期的那样,抑制辛德毕斯病毒(SINV)复制子的复制机制,IC50~1.94,~0.23,>1.25μM,分别。这些结果强调了重新利用药物作为针对CHIKV的快速和有效的抗病毒治疗剂的功效和希望。
Chikungunya 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 KD values ∼5.45 μM, ∼161.3 μM, and ∼3.83 μM, respectively. In a capillary electrophoresis-based assay, a decrease in CHIKV
nsP1 MTase activity was observed in a dose-dependent manner. Treatment with BH, ABT, and PT lead to a dose-dependent reduction in the virus titer with IC50 < 100, ∼6.75, 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 highlight the potential of repurposing drugs as rapid and effective antiviral therapeutics against CHIKV.