TY - JOUR
T1 - A small molecule compound with an indole moiety inhibits the main protease of SARS-CoV-2 and blocks virus replication
AU - Hattori, Shin ichiro
AU - Higashi-Kuwata, Nobuyo
AU - Hayashi, Hironori
AU - Allu, Srinivasa Rao
AU - Raghavaiah, Jakka
AU - Bulut, Haydar
AU - Das, Debananda
AU - Anson, Brandon J.
AU - Lendy, Emma K.
AU - Takamatsu, Yuki
AU - Takamune, Nobutoki
AU - Kishimoto, Naoki
AU - Murayama, Kazutaka
AU - Hasegawa, Kazuya
AU - Li, Mi
AU - Davis, David A.
AU - Kodama, Eiichi N.
AU - Yarchoan, Robert
AU - Wlodawer, Alexander
AU - Misumi, Shogo
AU - Mesecar, Andrew D.
AU - Ghosh, Arun K.
AU - Mitsuya, Hiroaki
N1 - Funding Information:
The authors thank Ms. Asuka Fujiwara, National Center for Global Health and Medicine (Tokyo, Japan), for technical help. The authors also thank Dr. Hiroyuki Hata, Kumamoto University (Kumamoto Japan), for helpful discussion. The present work was supported by a grant for Development of Novel Drugs for Treating COVID-19 from the Intramural Research Program of National Center for Global Health and Medicine (H.M., 19A3001 and S.H., 20A2001D), in part by Japan Agency for Medical Research and Development (AMED)(H.M., 20fk0108257), in part by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health (H.M. and A.W.), in part with Federal funds from the National Cancer Institute, NIH, under Contract No. HHSN261200800001E (to M.L.), and in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health (A.K.G., AI150466 and AI085089, and A.D.M., contract No. HHSN272201700060C). We also acknowledge the support of the Biomedical Research Core of the Tohoku University Graduate School of Medicine. The synchrotron radiation experiment was performed at BL41XU of SPring-8 with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (proposal no. 2020A1778). This study utilized the high-performance computational capabilities of the Biowulf Linux cluster at the National Institutes of Health, Bethesda, MD (https://hpc.nih.gov). The content of this publication is solely the responsibility of the authors and does not necessarily represent the official views or policies of the Department of Health and Human Services, nor does the mention of trade names, commercial products, or organizations imply endorsement by the U. S. Government.
Publisher Copyright:
© 2021, This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - Except remdesivir, no specific antivirals for SARS-CoV-2 infection are currently available. Here, we characterize two small-molecule-compounds, named GRL-1720 and 5h, containing an indoline and indole moiety, respectively, which target the SARS-CoV-2 main protease (Mpro). We use VeroE6 cell-based assays with RNA-qPCR, cytopathic assays, and immunocytochemistry and show both compounds to block the infectivity of SARS-CoV-2 with EC50 values of 15 ± 4 and 4.2 ± 0.7 μM for GRL-1720 and 5h, respectively. Remdesivir permitted viral breakthrough at high concentrations; however, compound 5h completely blocks SARS-CoV-2 infection in vitro without viral breakthrough or detectable cytotoxicity. Combination of 5h and remdesivir exhibits synergism against SARS-CoV-2. Additional X-ray structural analysis show that 5h forms a covalent bond with Mpro and makes polar interactions with multiple active site amino acid residues. The present data suggest that 5h might serve as a lead Mpro inhibitor for the development of therapeutics for SARS-CoV-2 infection.
AB - Except remdesivir, no specific antivirals for SARS-CoV-2 infection are currently available. Here, we characterize two small-molecule-compounds, named GRL-1720 and 5h, containing an indoline and indole moiety, respectively, which target the SARS-CoV-2 main protease (Mpro). We use VeroE6 cell-based assays with RNA-qPCR, cytopathic assays, and immunocytochemistry and show both compounds to block the infectivity of SARS-CoV-2 with EC50 values of 15 ± 4 and 4.2 ± 0.7 μM for GRL-1720 and 5h, respectively. Remdesivir permitted viral breakthrough at high concentrations; however, compound 5h completely blocks SARS-CoV-2 infection in vitro without viral breakthrough or detectable cytotoxicity. Combination of 5h and remdesivir exhibits synergism against SARS-CoV-2. Additional X-ray structural analysis show that 5h forms a covalent bond with Mpro and makes polar interactions with multiple active site amino acid residues. The present data suggest that 5h might serve as a lead Mpro inhibitor for the development of therapeutics for SARS-CoV-2 infection.
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U2 - 10.1038/s41467-021-20900-6
DO - 10.1038/s41467-021-20900-6
M3 - Article
C2 - 33510133
AN - SCOPUS:85100033738
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 668
ER -