Identification of a Direct-Acting Antiviral Agent Targeting RNA Helicase via a Graphene Oxide Nanobiosensor
Identification of a Direct-Acting Antiviral Agent Targeting RNA Helicase via a Graphene Oxide Nanobiosensor
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DOI：
10.1021/acsami.1c04641
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发表时间：
2021-05-26
影响因子：
9.5
通讯作者：
Min, Dal-Hee
中科院分区：
文献类型：
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作者：
Kim, Jungho;Park, Se-Jin;Min, Dal-Hee
登革病毒（DENV）是一种由蚊子传播的虫媒病毒，可引起登革热、登革出血热和登革休克综合征等传染病。尽管DENV构成危险，但没有批准的抗病毒药物用于治疗DENV感染。考虑到全球登革热爆发的可能性，快速开发针对DENV感染的抗病毒剂作为先发制人的措施至关重要;因此，建议选择明显的药物靶标，例如参与病毒生命周期的病毒酶。解旋酶是登革病毒中的潜在药物靶标，是一种重要的病毒酶，其在病毒复制期间解旋双链病毒RNA，释放单链RNA基因组。因此，解旋酶活性的抑制剂可以用作直接作用的抗病毒剂。在这里，我们介绍了一种基于氧化石墨烯的RNA解旋酶测定法，该方法能够基于荧光分析RNA底物特异性解旋酶的活性。该检测方法具有高可靠性和高通量筛选能力，可从FDA批准的药物库中鉴定新的解旋酶抑制剂候选物米卡芬净（MCFG）。MCFG作为靶向RNA解旋酶的直接作用抗病毒剂，抑制细胞和动物模型中的DENV增殖。值得注意的是，在体内，MCFG治疗减少了几种组织中的病毒血症、炎性细胞因子水平和病毒载量，并在致死小鼠模型中将存活率提高了40%。因此，我们建议MCFG作为一个潜在的直接作用的抗病毒药物候选人。
Dengue virus (DENV), an arbovirus transmitted by mosquitoes, causes infectious diseases such as dengue fever, dengue hemorrhagic fever, and dengue shock syndrome. Despite the dangers posed by DENV, there are no approved antiviral drugs for treatment of DENV infection. Considering the potential for a global dengue outbreak, rapid development of antiviral agents against DENV infections is crucial as a preemptive measure; thus, the selection of apparent drug targets, such as the viral enzymes involved in the viral life cycle, is recommended. Helicase, a potential drug target in DENV, is a crucial viral enzyme that unwinds double-stranded viral RNA, releasing single-stranded RNA genomes during viral replication. Therefore, an inhibitor of helicase activity could serve as a direct-acting antiviral agent. Here, we introduce an RNA helicase assay based on graphene oxide, which enables fluorescence-based analysis of RNA substrate-specific helicase enzyme activity. This assay demonstrated high reliability and ability for high-throughput screening, identifying a new helicase inhibitor candidate, micafungin (MCFG), from an FDA-approved drug library. As a direct-acting antiviral agent targeting RNA helicase, MCFG inhibits DENV proliferation in cells and an animal model. Notably, in vivo, MCFG treatment reduced viremia, inflammatory cytokine levels, and viral loads in several tissues and improved survival rates by up to 40% in a lethal mouse model. Therefore, we suggest MCFG as a potential direct-acting antiviral drug candidate.