目的 通過改變藥物與病毒引入的先后順序，探討氯喹與硫酸羥氯喹對(duì)嚴(yán)重急性呼吸綜合征冠狀病毒2（SARSCoV-2）不同變體（Prototype、Beta、Delta、Omicron）的體外抗病毒活性。方法 預(yù)防研究：Vero E6細(xì)胞用氯喹或硫酸羥氯喹（200.00、150.00、100.00、50.00、16.70、5.55、1.85、0.62、0.21μmol·L^-1）孵育1h，加入病毒附著2h，去除病毒-藥物混合物，用新鮮培養(yǎng)基培養(yǎng)細(xì)胞直到實(shí)驗(yàn)結(jié)束；治療研究：Vero E6細(xì)胞中加入病毒吸附2h，去除病毒，用含氯喹或硫酸羥氯喹的培養(yǎng)基培養(yǎng)細(xì)胞直到實(shí)驗(yàn)結(jié)束；全時(shí)研究：Vero E6細(xì)胞用藥物孵育1h，加入病毒附著2h，去除病毒-藥物混合物，用含藥物的培養(yǎng)基培養(yǎng)細(xì)胞直到實(shí)驗(yàn)結(jié)束。培養(yǎng)72h后顯微鏡下觀察細(xì)胞是否變圓脫落判斷細(xì)胞病變情況，計(jì)算半數(shù)效應(yīng)濃度（EC₅₀）及藥物選擇指數(shù)（SI）。結(jié)果 2種藥物對(duì)于SARS-CoV-2的預(yù)防效果較差；氯喹和硫酸羥氯喹在治療和全時(shí)處理下均表現(xiàn)出良好的抗病毒活性，其中硫酸羥氯喹的EC₅₀小于氯喹，SI大于氯喹，抗病毒效果較氯喹更優(yōu)越。在治療和全時(shí)處理?xiàng)l件下，氯喹（EC₅₀＝0.904μmol·L^-1）和硫酸羥氯喹（EC₅₀＝0.143μmol·L^-1）對(duì)Omicron變異株抗病毒效果較其他變種更明顯。結(jié)論 氯喹和硫酸羥氯喹在治療和全時(shí)處理下均表現(xiàn)出良好的抗病毒活性，且2種藥物對(duì)Omicron變異株的活性高于其他變種。

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

國(guó)家重點(diǎn)研發(fā)計(jì)劃“公共安全風(fēng)險(xiǎn)防控與應(yīng)急技術(shù)裝備”重點(diǎn)專項(xiàng)（2020YFC0841800）；國(guó)家重點(diǎn)研發(fā)計(jì)劃“公共安全風(fēng)險(xiǎn)防控與應(yīng)急技術(shù)裝備”重點(diǎn)專項(xiàng)（2020YFC0842100）；2021年省級(jí)科技創(chuàng)新專項(xiàng)立項(xiàng)項(xiàng)目（2021ACB005）