Identification of natural compounds with antiviral activities against SARS-associated coronavirus

Shi-You Li, Cong Chen, Hai-Qing Zhang, Hai-Yan Guo, Hui Wang, Lin Wang, Xiang Zhang, Shi-Neng Hua, Jun Yu, Pei-Gen Xiao, Rong-Song Li, Xuehai Tan, Shi-You Li, Cong Chen, Hai-Qing Zhang, Hai-Yan Guo, Hui Wang, Lin Wang, Xiang Zhang, Shi-Neng Hua, Jun Yu, Pei-Gen Xiao, Rong-Song Li, Xuehai Tan

Abstract

More than 200 Chinese medicinal herb extracts were screened for antiviral activities against Severe Acute Respiratory Syndrome-associated coronavirus (SARS-CoV) using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay for virus-induced cytopathic effect (CPE). Four of these extracts showed moderate to potent antiviral activities against SARS-CoV with 50% effective concentration (EC50) ranging from 2.4 +/- 0.2 to 88.2 +/- 7.7 microg/ml. Out of the four, Lycoris radiata was most potent. To identify the active component, L. radiata extract was subjected to further fractionation, purification, and CPE/MTS assays. This process led to the identification of a single substance lycorine as an anti-SARS-CoV component with an EC50 value of 15.7 +/- 1.2 nM. This compound has a CC50 value of 14980.0 +/- 912.0 nM in cytotoxicity assay and a selective index (SI) greater than 900. The results suggested that four herbal extracts and the compound lycorine are candidates for the development of new anti-SARS-CoV drugs in the treatment of SARS.

Figures

Fig. 1
Fig. 1
Effects of herb compound extracts on replication of SARS-CoV. The Vero E6 cell seeding, virus infection, compound addition, cell incubation, and measurement were described in the method. The percentage of CPE reduction was calculated by subtracting the mean of virus-infected cell control (0%) from the measured absorbance. The resulting number was divided by the uninfected cell control (100%). The mean values and the standard deviation (S.D.) are shown in the figures. Data presented are the average of duplicate values from three independent experiments. Magnification for visual observation: 200×.
Fig. 2
Fig. 2
Effect of fraction A, B, C, and D of L. radiata on inhibition of CPE caused by SARS-CoV in Vero E6 cells. OD value was measured in a CPE assay as described under experimental procedures. The percentage of CPE reduction was calculated by subtracting the mean of infected cell control (0%) from the measured absorbance. The resulting number was divided by the uninfected cell control (100%). The mean values and the standard deviation (S.D.) are shown in the figures. Data presented are the average of duplicate values from three independent experiments.
Fig. 3
Fig. 3
Chemical structure of lycorine.

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