The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines.Therefore,there remains a need for alternative antivirals tha...The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines.Therefore,there remains a need for alternative antivirals that target processes less susceptible to mutations,such as the formation of six-helix bundle(6-HB)during the viral fusion step of host cell entry.In this study,a novel high-throughput screening(HTS)assay employing a yeast-two-hybrid(Y2H)system was established to identify inhibitors of HR1/HR2 interaction.The compound IMB-9C,which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity,was selected.IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion.It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex.In addition,virtual docking and site-mutagenesis results suggest that amino acid residues A930,I931,K933,T941,and L945 are critical for IMB-9C binding to HR1.Collectively,in this study,we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.展开更多
基金supported by CAMS Innovation Fund for Medical Sciences(CIFMS)(Nos.2021-1-I2M-1-054,2022-I2M-2-002,2023-I2M-2-006,2020-I2M-2-010,and 2021-I2M-1-070,China)National Natural Science Foundation of China(No.81370087,China)+2 种基金Health and Medical Research Fund(HMRF)Commissioned Research on the Novel Coronavirus Disease(COVID1903010-Project 10,Hong Kong SAR,China)the PhD Research Startup Fund of Shanxi Medical University(XD2204,China)the Provincial Doctoral Fund of Shanxi Medical University(SD2312,China).
文摘The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines.Therefore,there remains a need for alternative antivirals that target processes less susceptible to mutations,such as the formation of six-helix bundle(6-HB)during the viral fusion step of host cell entry.In this study,a novel high-throughput screening(HTS)assay employing a yeast-two-hybrid(Y2H)system was established to identify inhibitors of HR1/HR2 interaction.The compound IMB-9C,which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity,was selected.IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion.It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex.In addition,virtual docking and site-mutagenesis results suggest that amino acid residues A930,I931,K933,T941,and L945 are critical for IMB-9C binding to HR1.Collectively,in this study,we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.
