Puhl Ana C, Fritch Ethan J, Lane Thomas R, Tse Longping V, Yount Boyd L, Sacramento Carolina Q, Fintelman-Rodrigues Natalia, Tavella Tatyana Almeida, Maranhão Costa Fabio Trindade, Weston Stuart, Logue James, Frieman Matthew, Premkumar Lakshmanane, Pearce Kenneth H, Hurst Brett L, Andrade Carolina Horta, Levi James A, Johnson Nicole J, Kisthardt Samantha C, Scholle Frank, Souza Thiago Moreno L, Moorman Nathaniel John, Baric Ralph S, Madrid Peter B, Ekins Sean
Collaborations Pharmaceuticals, Inc., 840 Main Campus Drive, Lab 3510, Raleigh, North Carolina 27606, United States.
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599, United States.
ACS Omega. 2021 Mar 10;6(11):7454-7468. doi: 10.1021/acsomega.0c05996. eCollection 2021 Mar 23.
Severe acute respiratory coronavirus 2 (SARS-CoV-2) is a newly identified virus that has resulted in over 2.5 million deaths globally and over 116 million cases globally in March, 2021. Small-molecule inhibitors that reverse disease severity have proven difficult to discover. One of the key approaches that has been widely applied in an effort to speed up the translation of drugs is drug repurposing. A few drugs have shown activity against Ebola viruses and demonstrated activity against SARS-CoV-2 . Most notably, the RNA polymerase targeting remdesivir demonstrated activity and efficacy in the early stage of the disease in humans. Testing other small-molecule drugs that are active against Ebola viruses (EBOVs) would appear a reasonable strategy to evaluate their potential for SARS-CoV-2. We have previously repurposed pyronaridine, tilorone, and quinacrine (from malaria, influenza, and antiprotozoal uses, respectively) as inhibitors of Ebola and Marburg viruses in HeLa cells and mouse-adapted EBOV in mice . We have now tested these three drugs in various cell lines (VeroE6, Vero76, Caco-2, Calu-3, A549-ACE2, HUH-7, and monocytes) infected with SARS-CoV-2 as well as other viruses (including MHV and HCoV 229E). The compilation of these results indicated considerable variability in antiviral activity observed across cell lines. We found that tilorone and pyronaridine inhibited the virus replication in A549-ACE2 cells with IC values of 180 nM and IC 198 nM, respectively. We used microscale thermophoresis to test the binding of these molecules to the spike protein, and tilorone and pyronaridine bind to the spike receptor binding domain protein with values of 339 and 647 nM, respectively. Human C for pyronaridine and quinacrine is greater than the IC observed in A549-ACE2 cells. We also provide novel insights into the mechanism of these compounds which is likely lysosomotropic.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是一种新发现的病毒，截至2021年3月，已在全球导致超过250万人死亡，全球确诊病例超过1.16亿例。事实证明，研发能逆转疾病严重程度的小分子抑制剂颇具难度。为加快药物转化进程，药物重新利用是一种广泛应用的关键方法。一些药物已显示出对埃博拉病毒有活性，并证明对SARS-CoV-2也有活性。最值得注意的是，靶向瑞德西韦的RNA聚合酶在人类疾病早期显示出活性和疗效。测试其他对埃博拉病毒（EBOV）有活性的小分子药物似乎是评估其对SARS-CoV-2潜在作用的合理策略。我们之前已将咯萘啶、替洛隆和奎纳克林（分别来自疟疾、流感和抗原生动物用途）重新用作HeLa细胞中埃博拉病毒和马尔堡病毒以及小鼠适应性EBOV的抑制剂。我们现在已在感染SARS-CoV-2以及其他病毒（包括鼠肝炎病毒和人冠状病毒229E）的各种细胞系（VeroE6、Vero76、Caco-2、Calu-3、A549-ACE2、HUH-7和单核细胞）中测试了这三种药物。这些结果汇总表明，各细胞系中观察到的抗病毒活性存在很大差异。我们发现替洛隆和咯萘啶分别以180 nM和198 nM的IC值抑制A549-ACE2细胞中的病毒复制。我们使用微量热泳动法测试这些分子与刺突蛋白的结合，替洛隆和咯萘啶分别以339 nM和647 nM的Kd值与刺突受体结合域蛋白结合。咯萘啶和奎纳克林的人体Cmax大于在A549-ACE2细胞中观察到的IC值。我们还对这些化合物的作用机制提供了新见解，其作用机制可能是亲溶酶体性的。
