A Comparative Analysis of SARS-CoV-2 Antivirals Characterizes 3CLpro Inhibitor PF-00835231 as a Potential New Treatment for COVID-19
A Comparative Analysis of SARS-CoV-2 Antivirals Characterizes 3CLpro Inhibitor PF-00835231 as a Potential New Treatment for COVID-19
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DOI：
10.1128/jvi.01819-20
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发表时间：
2021-05-01
影响因子：
5.4
通讯作者：
Dittmann, Meike
中科院分区：
文献类型：
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作者：
de Vries, Maren;Mohamed, Adil S.;Dittmann, Meike
严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)是冠状病毒病2019(新冠肺炎)的病原体。迫切需要新的有效的抗病毒药物来治疗新冠肺炎，因为到目前为止唯一被批准的直接作用的抗病毒药物是redesivir，靶向病毒聚合酶复合体。病毒生命周期中一个潜在的替代靶点是主要的SARS-CoV-2蛋白酶3CL(Pro)(M-Pro)。候选药物PF-00835231是第一个临床试验中的抗3CL(PRO)方案的活性化合物。在这里，我们进行了一个比较分析的PF-00835231，临床前3CL(PRO)抑制剂GC-376，和聚合酶抑制剂雷米西韦，在修饰表达血管紧张素转换酶2的肺泡基底上皮细胞(A549(+血管紧张素转换酶2)细胞)。我们发现PF-00835231的效价至少与瑞维韦或GC-376相似或更高。药物加药时间方法描述了A549(+ACE2)细胞中SARS-CoV-2早期生命周期步骤的时间，并验证了PF-00835231‘S的早期作用时间。在一个人体极化的呼吸道上皮细胞模型中，PF-00835231和雷米昔韦在低微摩尔浓度下都能有效地抑制SARS-CoV-2。最后，我们发现，外排转运蛋白P-糖蛋白在A549(+ACE2)细胞和人极化呼吸道上皮细胞培养中都不会对PF-00835231‘S的疗效产生负面影响。因此，我们的研究为PF-00835231作为一种有效的SARS-CoV-2抗病毒药物的潜力提供了体外证据，并解决了基于先前在非人类体外模型中出现的担忧。目前只有一种直接作用的抗病毒药物获得批准，即病毒聚合酶抑制剂瑞德韦，其疗效有限。因此，有必要开发更多的抗病毒化合物，使其副作用最小，并可替代病毒靶点。其中一个替代靶点是其主要的蛋白酶3CL(Pro)(M-Pro)，这是SARS-CoV-2生命周期的重要组成部分，将病毒多蛋白加工成病毒聚合酶复合体的组成部分。在这项研究中，我们描述了一种新型抗病毒药物PF-00835231，它是临床试验中第一类3CL(PRO)靶向方案的活性成分。使用人呼吸道上皮细胞的3D体外模型，我们展示了PF-00835231抑制SARS-CoV-2的抗病毒潜力。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of coronavirus disease 2019 (COVID-19). There is a dire need for novel effective antivirals to treat COVID-19, as the only approved direct-acting antiviral to date is remdesivir, targeting the viral polymerase complex. A potential alternate target in the viral life cycle is the main SARS-CoV-2 protease 3CL(pro) (M-pro). The drug candidate PF-00835231 is the active compound of the first anti-3CL(pro) regimen in clinical trials. Here, we perform a comparative analysis of PF-00835231, the preclinical 3CL(pro) inhibitor GC-376, and the polymerase inhibitor remdesivir, in alveolar basal epithelial cells modified to express ACE2 (A549(+ACE2) cells). We find PF-00835231 with at least similar or higher potency than remdesivir or GC-376. A time-of-drug-addition approach delineates the timing of early SARS-CoV-2 life cycle steps in A549(+ACE2) cells and validates PF-00835231's early time of action. In a model of the human polarized airway epithelium, both PF-00835231 and remdesivir potently inhibit SARS-CoV-2 at low micromolar concentrations. Finally, we show that the efflux transporter P-glycoprotein, which was previously suggested to diminish PF-00835231's efficacy based on experiments in monkey kidney Vero E6 cells, does not negatively impact PF-00835231 efficacy in either A549(+ACE2) cells or human polarized airway epithelial cultures. Thus, our study provides in vitro evidence for the potential of PF-00835231 as an effective SARS-CoV-2 antiviral and addresses concerns that emerged based on prior studies in nonhuman in vitro models.IMPORTANCE The arsenal of SARS-CoV-2 specific antiviral drugs is extremely limited. Only one direct-acting antiviral drug is currently approved, the viral polymerase inhibitor remdesivir, and it has limited efficacy. Thus, there is a substantial need to develop additional antiviral compounds with minimal side effects and alternate viral targets. One such alternate target is its main protease, 3CL(pro) (M-pro), an essential component of the SARS-CoV-2 life cycle processing the viral polyprotein into the components of the viral polymerase complex. In this study, we characterize a novel antiviral drug, PF-00835231, which is the active component of the first-in-class 3CL(pro)-targeting regimen in clinical trials. Using 3D in vitro models of the human airway epithelium, we demonstrate the antiviral potential of PF-00835231 for inhibition of SARS-CoV-2.