A library of nucleotide analogues terminate RNA synthesis catalyzed by polymerases of coronaviruses that cause SARS and COVID-19
A library of nucleotide analogues terminate RNA synthesis catalyzed by polymerases of coronaviruses that cause SARS and COVID-19
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DOI：
10.1016/j.antiviral.2020.104857
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发表时间：
2020-08-01
影响因子：
7.6
通讯作者：
Ju, Jingyue
中科院分区：
文献类型：
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作者：
Jockusch, Steffen;Tao, Chuanjuan;Ju, Jingyue
SARS-CoV-2是冠状病毒家族的一员，是导致当前COVID-19全球大流行的原因。我们先前证明了五种核苷酸类似物抑制SARS-CoV-2 RNA依赖性RNA聚合酶（RdRp），包括Sofosbuvir，Alovudine，Zidovudine，Tenofovir alafenamide和Emtricitabine的活性三磷酸形式。我们在这里报告的评价与各种结构和化学特征的核苷三磷酸类似物作为抑制剂的RdRps的SARS-CoV和SARS-CoV-2的库。这些特征包括糖上或碱基上的修饰（2'或3'修饰、碳环、无环或双脱氧核苷酸）。目标是鉴定出不仅能终止这些冠状病毒RdRps催化的RNA合成，而且还具有抵抗病毒外切核酸酶活性的潜力的核苷酸类似物。我们检查了这些核苷酸类似物在聚合酶反应中被RdRps掺入并防止进一步掺入的能力。虽然所有11种检测分子均显示掺入，但6种显示聚合酶反应立即终止（卡波韦、更昔洛韦、司他夫定和恩替卡韦的三磷酸盐; 3 '-OMe-UTP和生物素-16-dUTP），2种显示延迟终止（西多福韦二磷酸盐和2'-OMe-UTP），3种未终止聚合酶反应（2 '-F-dUTP、2'-NH 2-dUTP和脱硫生物素-16-UTP）。冠状病毒拥有一种核酸外切酶，显然需要在生长的RNA链的3 '-末端的2'-OH进行校对。在这项研究中，所有核苷三磷酸类似物评估形成沃森-克里克样碱基对。显示终止的核苷酸类似物缺乏2 '-OH，具有封闭的2'-OH，或显示延迟的终止。因此，这些核苷酸类似物对于进一步研究以评估它们是否可以逃避病毒核酸外切酶活性是有意义的。这些核苷酸类似物中的五种（西多福韦、阿巴卡韦、缬更昔洛韦/更昔洛韦、司他夫定和恩替卡韦）的前药是FDA批准的用于治疗其他病毒感染的药物，并且它们的安全性特征已经得到充分确立。在细胞培养中证明抑制病毒复制的效力后，候选分子可以作为COVID-19的潜在疗法进行快速评估。
SARS-CoV-2, a member of the coronavirus family, is responsible for the current COVID-19 worldwide pandemic. We previously demonstrated that five nucleotide analogues inhibit the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp), including the active triphosphate forms of Sofosbuvir, Alovudine, Zidovudine, Tenofovir alafenamide and Emtricitabine. We report here the evaluation of a library of nucleoside triphosphate analogues with a variety of structural and chemical features as inhibitors of the RdRps of SARS-CoV and SARS-CoV-2. These features include modifications on the sugar (2' or 3' modifications, carbocyclic, acyclic, or dideoxynucleotides) or on the base. The goal is to identify nucleotide analogues that not only terminate RNA synthesis catalyzed by these coronavirus RdRps, but also have the potential to resist the viruses' exonuclease activity. We examined these nucleotide analogues for their ability to be incorporated by the RdRps in the polymerase reaction and to prevent further incorporation. While all 11 molecules tested displayed incorporation, 6 exhibited immediate termination of the polymerase reaction (triphosphates of Carbovir, Ganciclovir, Stavudine and Entecavir; 3'-OMe-UTP and Biotin-16-dUTP), 2 showed delayed termination (Cidofovir diphosphate and 2'-OMe-UTP), and 3 did not terminate the polymerase reaction (2'-F-dUTP, 2'-NH2-dUTP and Desthiobiotin-16-UTP). The coronaviruses possess an exonuclease that apparently requires a 2'-OH at the 3'-terminus of the growing RNA strand for proofreading. In this study, all nucleoside triphosphate analogues evaluated form Watson-Crick-like base pairs. The nucleotide analogues demonstrating termination either lack a 2'-OH, have a blocked 2'-OH, or show delayed termination. Thus, these nucleotide analogues are of interest for further investigation to evaluate whether they can evade the viral exonuclease activity. Prodrugs of five of these nucleotide analogues (Cidofovir, Abacavir, Valganciclovir/Ganciclovir, Stavudine and Entecavir) are FDA-approved medications for treatment of other viral infections, and their safety profiles are well established. After demonstrating potency in inhibiting viral replication in cell culture, candidate molecules can be rapidly evaluated as potential therapies for COVID-19.