Identification of small molecule inhibitors of the Chikungunya virus nsP1 RNA capping enzyme.
Identification of small molecule inhibitors of the Chikungunya virus nsP1 RNA capping enzyme.
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DOI：
10.1016/j.antiviral.2018.03.013
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发表时间：
2018-06
影响因子：
7.6
通讯作者：
Geiss BJ
中科院分区：
文献类型：
--
作者：
Feibelman KM;Fuller BP;Li L;LaBarbera DV;Geiss BJ
基孔肯雅病毒（CHIKV）是一种节肢动物传播的甲病毒。甲病毒是一种正链RNA病毒，它需要一个5´cap结构来直接翻译病毒多蛋白，并防止宿主细胞核酸酶降解病毒RNA基因组。5´RNA帽的形成是由病毒蛋白nsP1策划的，该蛋白结合GTP并提供帽形成所必需的N-7甲基转移酶和鸟苷基转移酶活性。具有异常nsP1活性的病毒无法有效复制，这表明nsP1是发现抗病毒药物的一个有希望的靶标。鉴于市面上没有针对CHIKV的抗病毒治疗方法，必须确定可开发为潜在治疗方法的化合物。本研究详细介绍了利用基于荧光偏振的GTP竞争分析法，从含有fda批准的抗CHIKV nsP1药物、天然产物和已知生物活性的文库中筛选出3051种化合物的高通量筛选。在低摩尔浓度下，来自该筛选的几个小分子能够与GTP竞争CHIKV nsP1 GTP结合位点。用正交试验对化合物进行了评价，该试验测量了nsP1在抑制剂存在的情况下进行盖顶反应的鸟酰化步骤的能力。此外，用CHIKV病毒和密切相关的甲病毒Sindbis病毒的活病毒试验与细胞毒性试验结合使用，以确定细胞培养物中化合物的抗病毒活性。在体外实验中发现，天然衍生的化合物四巴比酸可以抑制CHIKV nsP1 GTP结合和鸟苷化，并在24 hpi和48 hpi条件下减弱病毒在鼠BHK21和人Huh 7细胞系中的生长。这些数据表明，叶酸的开发和进一步探索CHIKV nsP1作为药物靶点可能有助于抗甲型病毒药物开发策略的进展。
Chikungunya virus (CHIKV) is an arthropod-borne alphavirus. Alphaviruses are positive strand RNA viruses that require a 5´ cap structure to direct translation of the viral polyprotein and prevent degradation of the viral RNA genome by host cell nucleases. Formation of the 5´ RNA cap is orchestrated by the viral protein nsP1, which binds GTP and provides the N-7 methyltransferase and guanylyltransferase activities that are necessary for cap formation. Viruses with aberrant nsP1 activity are unable to replicate effectively suggesting that nsP1 is a promising target for antiviral drug discovery. Given the absence of commercially available antiviral therapies for CHIKV, it is imperative to identify compounds that could be developed as potential therapeutics. This study details a high-throughput screen of 3051 compounds from libraries containing FDA-approved drugs, natural products, and known bioactives against CHIKV nsP1 using a fluorescence polarization-based GTP competition assay. Several small molecule hits from this screen were able to compete with GTP for the CHIKV nsP1 GTP binding site at low molar concentrations. Compounds were also evaluated with an orthogonal assay that measured the ability of nsP1 to perform the guanylation step of the capping reaction in the presence of inhibitor. In addition, live virus assays with CHIKV and closely related alphavirus, Sindbis virus, were used in conjunction with cell toxicity assays to determine the antiviral activity of compounds in cell culture. The naturally derived compound lobaric acid was found to inhibit CHIKV nsP1 GTP binding and guanylation as well as attenuate viral growth in vitro at both 24 hpi and 48 hpi in hamster BHK21 and human Huh 7 cell lines. These data indicate that development of lobaric acid and further exploration of CHIKV nsP1 as a drug target may aid in the progress of anti-alphaviral drug development strategies.
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影响因子：
5.6
作者：
Geiss, Brian J.;Thompson, Aaron A.;Andrews, Andrew J.;Sons, Robert L.;Gari, Hamid H.;Keenan, Susan M.;Peersen, Olve B.
通讯作者：
Peersen, Olve B.
影响因子：
7.6
作者：
Gigante, Alba;Gomez-SanJuan, Asier;Perez-Perez, Maria-Jesus
通讯作者：
Perez-Perez, Maria-Jesus
影响因子：
3.1
作者：
Jose J;Snyder JE;Kuhn RJ
通讯作者：
Kuhn RJ
影响因子：
4.8
作者：
Laakkonen, P;Ahola, T;Kaariainen, L
通讯作者：
Kaariainen, L
影响因子：
11.8
作者：
Borgherini, Gianandrea;Poubeau, Patrice;Paganin, Fabrice
通讯作者：
Paganin, Fabrice