The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner
The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner
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DOI：
10.3390/v12060629
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发表时间：
2020-06-01
期刊：
影响因子：
4.7
通讯作者：
Inoue, Jun-ichiro
中科院分区：
文献类型：
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作者：
Yamamoto, Mizuki;Kiso, Maki;Inoue, Jun-ichiro
尽管冠状病毒肺炎 (COVID-19) 的病原体 SARS-CoV-2 感染正在全球范围内迅速传播，但尚未有药物被证明能够充分有效地治疗 COVID-19。我们之前发现甲磺酸萘莫司他是一种用于弥散性血管内凝血（DIC）的现有药物，通过靶向跨膜丝氨酸蛋白酶2（TMPRSS2）有效阻断中东呼吸综合征冠状病毒（MERS-CoV）S蛋白介导的细胞融合，并抑制MERS-CoV对人肺上皮来源的Calu-3细胞的感染。在此，我们建立了一种依赖于严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) S 蛋白、血管紧张素 I 转换酶 2 (ACE2) 和 TMPRSS2 的定量融合测定，发现甲磺酸萘莫司他有效抑制融合，而甲磺酸卡莫司他的活性大约低 10 倍。此外，甲磺酸萘莫司他阻断Calu-3细胞的SARS-CoV-2感染，有效浓度（EC）（50）约为10 nM，低于连续输注静脉给药后的平均血药浓度。另一方面，VeroE6/TMPRSS2细胞需要明显更高的剂量（EC(50)约30μM），其中TMPRSS2独立但组织蛋白酶依赖性内体感染途径可能占主导地位。总之，我们的研究表明，nafamostat mesylate 可在细胞融合测定系统中有效抑制 SARS-CoV-2 S 蛋白介导的融合，并以细胞类型依赖性方式在体外抑制 SARS-CoV-2 感染。这些发现加上有关 nafamostat 安全性的临床数据，使其成为治疗 COVID-19 的可能候选药物。
Although infection by SARS-CoV-2, the causative agent of coronavirus pneumonia disease (COVID-19), is spreading rapidly worldwide, no drug has been shown to be sufficiently effective for treating COVID-19. We previously found that nafamostat mesylate, an existing drug used for disseminated intravascular coagulation (DIC), effectively blocked Middle East respiratory syndrome coronavirus (MERS-CoV) S protein-mediated cell fusion by targeting transmembrane serine protease 2 (TMPRSS2), and inhibited MERS-CoV infection of human lung epithelium-derived Calu-3 cells. Here we established a quantitative fusion assay dependent on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) S protein, angiotensin I converting enzyme 2 (ACE2) and TMPRSS2, and found that nafamostat mesylate potently inhibited the fusion while camostat mesylate was about 10-fold less active. Furthermore, nafamostat mesylate blocked SARS-CoV-2 infection of Calu-3 cells with an effective concentration (EC)(50)around 10 nM, which is below its average blood concentration after intravenous administration through continuous infusion. On the other hand, a significantly higher dose (EC(50)around 30 mu M) was required for VeroE6/TMPRSS2 cells, where the TMPRSS2-independent but cathepsin-dependent endosomal infection pathway likely predominates. Together, our study shows that nafamostat mesylate potently inhibits SARS-CoV-2 S protein-mediated fusion in a cell fusion assay system and also inhibits SARS-CoV-2 infection in vitro in a cell-type-dependent manner. These findings, together with accumulated clinical data regarding nafamostat's safety, make it a likely candidate drug to treat COVID-19.