Jin Young-Hee, Jeon Sangeun, Lee Jihye, Kim Seungtaek, Jang Min Seong, Park Chul Min, Song Jong Hwan, Kim Hyoung Rae, Kwon Sunoh
KM Application Center, Korea Institute of Oriental Medicine, Daegu 41062, Korea.
Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology, Daejeon 34114, Korea.
Pharmaceutics. 2021 Nov 2;13(11):1839. doi: 10.3390/pharmaceutics13111839.
Cardiotonic steroids are steroid-like natural compounds known to inhibit Na/K-ATPase pumps. To develop a broad-spectrum antiviral drug against the emerging coronavirus infection, this study assessed the antiviral properties of these compounds. The activity of seven types of cardiotonic steroids against the MERS-CoV, SARS-CoV, and SARS-CoV-2 coronavirus varieties was analyzed using immunofluorescence antiviral assay in virus-infected cells. Bufalin, cinobufagin, and telocinobufagin showed high anti-MERS-CoV activities (IC, 0.0170.027 μM); bufalin showed the most potent anti-SARS-CoV and SARS-CoV-2 activity (IC, 0.0160.019 μM); cinobufotalin and resibufogenin showed comparatively low anti-coronavirus activity (IC, 0.231~1.612 μM). Differentially expressed genes in Calu3 cells treated with cinobufagin, telocinobufagin, or bufalin, which had high antiviral activity during MERS-CoV infection were analyzed using QuantSeq 3' mRNA-Seq analysis and data showed similar gene expression patterns. Furthermore, the intraperitoneal administration of 10 mg/kg/day bufalin, cinobufagin, or digitoxin induced 100% death after 1, 2, and 4 days in 5-day repeated dose toxicity studies and it indicated that bufalin had the strongest toxicity. Pharmacokinetic studies suggested that telocinobufagin, which had high anti-coronavirus activity and low toxicity, had better microsomal stability, lower CYP inhibition, and better oral bioavailability than cinobufagin. Therefore, telocinobufagin might be the most promising cardiotonic steroid as a therapeutic for emerging coronavirus infections, including COVID-19.
强心甾类是一类已知可抑制钠钾ATP酶泵的类固醇样天然化合物。为开发一种针对新型冠状病毒感染的广谱抗病毒药物，本研究评估了这些化合物的抗病毒特性。使用免疫荧光抗病毒试验在病毒感染细胞中分析了七种强心甾类对中东呼吸综合征冠状病毒（MERS-CoV）、严重急性呼吸综合征冠状病毒（SARS-CoV）和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）变种的活性。蟾毒灵、华蟾酥毒基和远华蟾毒基表现出高抗MERS-CoV活性（半数抑制浓度[IC]，0.0170.027μM）；蟾毒灵表现出最强的抗SARS-CoV和SARS-CoV-2活性（IC，0.0160.019μM）；脂蟾毒配基和脂蟾毒精表现出相对较低的抗冠状病毒活性（IC，0.231~1.612μM）。使用QuantSeq 3' mRNA测序分析对在MERS-CoV感染期间具有高抗病毒活性的经华蟾酥毒基、远华蟾毒基或蟾毒灵处理的Calu3细胞中的差异表达基因进行分析，数据显示出相似的基因表达模式。此外，在为期5天的重复剂量毒性研究中，腹腔注射10mg/kg/天的蟾毒灵、华蟾酥毒基或洋地黄毒苷在1、2和4天后导致100%死亡，这表明蟾毒灵毒性最强。药代动力学研究表明，具有高抗冠状病毒活性和低毒性的远华蟾毒基比华蟾酥毒基具有更好的微粒体稳定性、更低的细胞色素P450（CYP）抑制作用和更好的口服生物利用度。因此，远华蟾毒基可能是作为包括2019冠状病毒病（COVID-19）在内的新型冠状病毒感染治疗药物最有前景的强心甾类。
