目的 通过改变药物与病毒引入的先后顺序，探讨氯喹与硫酸羟氯喹对严重急性呼吸综合征冠状病毒2（SARSCoV-2）不同变体（Prototype、Beta、Delta、Omicron）的体外抗病毒活性。方法 预防研究：Vero E6细胞用氯喹或硫酸羟氯喹（200.00、150.00、100.00、50.00、16.70、5.55、1.85、0.62、0.21μmol·L^-1）孵育1h，加入病毒附着2h，去除病毒-药物混合物，用新鲜培养基培养细胞直到实验结束；治疗研究：Vero E6细胞中加入病毒吸附2h，去除病毒，用含氯喹或硫酸羟氯喹的培养基培养细胞直到实验结束；全时研究：Vero E6细胞用药物孵育1h，加入病毒附着2h，去除病毒-药物混合物，用含药物的培养基培养细胞直到实验结束。培养72h后显微镜下观察细胞是否变圆脱落判断细胞病变情况，计算半数效应浓度（EC₅₀）及药物选择指数（SI）。结果 2种药物对于SARS-CoV-2的预防效果较差；氯喹和硫酸羟氯喹在治疗和全时处理下均表现出良好的抗病毒活性，其中硫酸羟氯喹的EC₅₀小于氯喹，SI大于氯喹，抗病毒效果较氯喹更优越。在治疗和全时处理条件下，氯喹（EC₅₀＝0.904μmol·L^-1）和硫酸羟氯喹（EC₅₀＝0.143μmol·L^-1）对Omicron变异株抗病毒效果较其他变种更明显。结论 氯喹和硫酸羟氯喹在治疗和全时处理下均表现出良好的抗病毒活性，且2种药物对Omicron变异株的活性高于其他变种。

Objective To investigated the in vitro antiviral activity of chloroquine and hydroxychloroquine sulfate against different variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (Prototype, Beta, Delta, Omicron) by changing the sequence of drug and virus introduction. Methods Prophylactic treatment: Vero E6 cells were treated with Chloroquine or hydroxychloroquine sulfate (200.00, 150.00, 100.00, 50.00, 16.70, 5.55, 1.85, 0.62, 0.21 μmol·L^-1) for 1h, then the virus was added and incubated for another 2 h. The virus-drug mixture was repalced with fresh medium until the end of the experiment. Post-entry treatment: Vero E6 cells were incubated with virus for 2 h, then the virus was removed and the cells were cultured with drugcontaining medium until the end of the experiment. Full-time treatment: Vero E6 cells were pretreated with the drug for 1h ahead, then virus was added and incubated for another 2 h. The virus-drug mixture was discarded and the cells were cultured with drugcontaining medium until the end of the experiment. After 72h of culture, the cells were observed to see whether they became round and shed to determine the cytopathic situation, and the semi-maximum effect concentration (EC₅₀) and drug selection index (SI) were calculated. Results Both drugs were less effective in preventing SARS-CoV-2. Chloroquine/hydroxychloroquine sulfate showed good antiviral activity under both therapeutic and full-time treatment. EC₅₀ of hydroxychloroquine sulfate was less than chloroquine, SI was greater than chloroquine, antiviral effect of hydroxychloroquine sulfate was better than chloroquine. The antiviral effect of chloroquine (EC₅₀ = 0.904 μmol·L^-1) and hydroxychloroquine sulfate (EC₅₀ = 0.143 μmol·L^-1) was more significant against Omicron variant than other variants under therapeutic and full-time treatment conditions. Conclusion Chloroquine/hydroxychloroquine sulfate showed good antiviral activity under both therapeutic and full-time treatment, and both drugs were significantly more active against the Omicron variant than the other variants.

R978.7

国家重点研发计划“公共安全风险防控与应急技术装备”重点专项（2020YFC0841800）；国家重点研发计划“公共安全风险防控与应急技术装备”重点专项（2020YFC0842100）；2021年省级科技创新专项立项项目（2021ACB005）