Identification of Z-Tyr-Ala-CHN
        2, a Cathepsin L Inhibitor with Broad-Spectrum Cell-Specific Activity against Coronaviruses, including SARS-CoV-2.

Jordi Doijen; Koen Temmerman; Christel Van den Eynde; Annick Diels; Nick Van den Broeck; Michiel Van Gool; Inha Heo; Steffen Jaensch; Marleen Zwaagstra; Mayra Diosa Toro; Winston Chiu; Steven De Jonghe; Pieter Leyssen; Denisa Bojkova; Sandra Ciesek; Jindrich Cinatl; Lore Verschueren; Christophe Buyck; Frank Van Kuppeveld; Johan Neyts; Marnix Van Loock; Ellen Van Damme

doi:10.3390/microorganisms11030717

Identification of Z-Tyr-Ala-CHN 2, a Cathepsin L Inhibitor with Broad-Spectrum Cell-Specific Activity against Coronaviruses, including SARS-CoV-2.

Jordi Doijen, Koen Temmerman, Christel Van den Eynde, Annick Diels, Nick Van den Broeck, Michiel Van Gool, Inha Heo, Steffen Jaensch, Marleen Zwaagstra, Mayra Diosa Toro, Winston Chiu, Steven De Jonghe, Pieter Leyssen, Denisa Bojkova, Sandra Ciesek, Jindrich Cinatl, Lore Verschueren, Christophe Buyck, Frank Van Kuppeveld, Johan NeytsMarnix Van Loock, Ellen Van Damme^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The ongoing COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is partly under control by vaccination. However, highly potent and safe antiviral drugs for SARS-CoV-2 are still needed to avoid development of severe COVID-19. We report the discovery of a small molecule, Z-Tyr-Ala-CHN 2, which was identified in a cell-based antiviral screen. The molecule exerts sub-micromolar antiviral activity against SARS-CoV-2, SARS-CoV-1, and human coronavirus 229E. Time-of-addition studies reveal that Z-Tyr-Ala-CHN 2 acts at the early phase of the infection cycle, which is in line with the observation that the molecule inhibits cathepsin L. This results in antiviral activity against SARS-CoV-2 in VeroE6, A549-hACE2, and HeLa-hACE2 cells, but not in Caco-2 cells or primary human nasal epithelial cells since the latter two cell types also permit entry via transmembrane protease serine subtype 2 (TMPRSS2). Given their cell-specific activity, cathepsin L inhibitors still need to prove their value in the clinic; nevertheless, the activity profile of Z-Tyr-Ala-CHN 2 makes it an interesting tool compound for studying the biology of coronavirus entry and replication.

Original language	English
Article number	717
Number of pages	20
Journal	Microorganisms
Volume	11
Issue number	3
DOIs	https://doi.org/10.3390/microorganisms11030717
Publication status	Published - 10 Mar 2023

Bibliographical note

Publisher Copyright:
© 2023 by the authors.

Funding

This study was supported by Janssen Research & Development, LLC, and has been executed as part of the Corona Accelerated R&D in Europe (CARE) project under grant agreement number 101005077 with the Innovative Medicines Initiative 2 Joint Undertaking (JU). The JU receives support from the European Union’s Horizon 2020 research and innovation program, EFPIA, Bill & Melinda Gates Foundation, Global Health Drug Discovery Institute, and the University of Dundee. This project has also been funded in part with federal funds from the Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority (BARDA), under OTA numbers HHSO100201700018C and HHSO100201800012C. The SARS-CoV-2 screening in VeroE6-eGFP cells was performed using the ‘Caps-It’ research infrastructure (project ZW13–02) that was financially supported by the Hercules Foundation (Research Foundation Flanders) and Rega Foundation, KU Leuven.

Funders	Funder number
Global Health Drug Discovery Institute
Office of the Assistant Secretary for Preparedness and Response
Bill and Melinda Gates Foundation
University of Dundee
Horizon 2020 Framework Programme
Biomedical Advanced Research and Development Authority	HHSO100201800012C, HHSO100201700018C, ZW13–02
European Federation of Pharmaceutical Industries and Associations
Fonds Wetenschappelijk Onderzoek
KU Leuven
Herculesstichting
Rega Foundation

Keywords

cathepsin L inhibitor
coronavirus
in vitro
phenotypic screening
SARS-CoV-2

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.3390/microorganisms11030717Licence: CC BY

microorganisms-11-00717-v2Final published version, 3.48 MBLicence: CC BY

Cite this

Doijen, J., Temmerman, K., Van den Eynde, C., Diels, A., Van den Broeck, N., Van Gool, M., Heo, I., Jaensch, S., Zwaagstra, M., Diosa Toro, M., Chiu, W., De Jonghe, S., Leyssen, P., Bojkova, D., Ciesek, S., Cinatl, J., Verschueren, L., Buyck, C., Van Kuppeveld, F., ... Van Damme, E. (2023). Identification of Z-Tyr-Ala-CHN 2, a Cathepsin L Inhibitor with Broad-Spectrum Cell-Specific Activity against Coronaviruses, including SARS-CoV-2. Microorganisms, 11(3), Article 717. https://doi.org/10.3390/microorganisms11030717

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abstract = "The ongoing COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is partly under control by vaccination. However, highly potent and safe antiviral drugs for SARS-CoV-2 are still needed to avoid development of severe COVID-19. We report the discovery of a small molecule, Z-Tyr-Ala-CHN 2, which was identified in a cell-based antiviral screen. The molecule exerts sub-micromolar antiviral activity against SARS-CoV-2, SARS-CoV-1, and human coronavirus 229E. Time-of-addition studies reveal that Z-Tyr-Ala-CHN 2 acts at the early phase of the infection cycle, which is in line with the observation that the molecule inhibits cathepsin L. This results in antiviral activity against SARS-CoV-2 in VeroE6, A549-hACE2, and HeLa-hACE2 cells, but not in Caco-2 cells or primary human nasal epithelial cells since the latter two cell types also permit entry via transmembrane protease serine subtype 2 (TMPRSS2). Given their cell-specific activity, cathepsin L inhibitors still need to prove their value in the clinic; nevertheless, the activity profile of Z-Tyr-Ala-CHN 2 makes it an interesting tool compound for studying the biology of coronavirus entry and replication. ",

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Doijen, J, Temmerman, K, Van den Eynde, C, Diels, A, Van den Broeck, N, Van Gool, M, Heo, I, Jaensch, S, Zwaagstra, M, Diosa Toro, M, Chiu, W, De Jonghe, S, Leyssen, P, Bojkova, D, Ciesek, S, Cinatl, J, Verschueren, L, Buyck, C, Van Kuppeveld, F, Neyts, J, Van Loock, M & Van Damme, E 2023, 'Identification of Z-Tyr-Ala-CHN 2, a Cathepsin L Inhibitor with Broad-Spectrum Cell-Specific Activity against Coronaviruses, including SARS-CoV-2.', Microorganisms, vol. 11, no. 3, 717. https://doi.org/10.3390/microorganisms11030717

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AU - Van den Eynde, Christel

AU - Diels, Annick

AU - Van den Broeck, Nick

AU - Van Gool, Michiel

AU - Heo, Inha

AU - Jaensch, Steffen

AU - Zwaagstra, Marleen

AU - Diosa Toro, Mayra

AU - Chiu, Winston

AU - De Jonghe, Steven

AU - Leyssen, Pieter

AU - Bojkova, Denisa

AU - Ciesek, Sandra

AU - Cinatl, Jindrich

AU - Verschueren, Lore

AU - Buyck, Christophe

AU - Van Kuppeveld, Frank

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