Methodological development of a multi-readout assay for the assessment of antiviral drugs against sars-cov-2

Hahn F, Häge S, Herrmann A, Wangen C, Kicuntod J, Jungnickl D, Tillmanns J, Müller R, Fraedrich K, Überla K, Kohlhof H, Enßer A, Marschall M (2021)

Publication Type: Journal article

Publication year: 2021

Journal

Pathogens MDPI

Book Volume: 10

Article Number: 1076

Journal Issue: 9

DOI: 10.3390/pathogens10091076

Abstract

Currently, human infections with the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) are accelerating the ongoing spread of the pandemic. Several innovative types of vaccines have already been developed, whereas effective options of antiviral treatments still await a scientific implementation. The development of novel anti-SARS-CoV-2 drug candidates demands skillful strategies and analysis systems. Promising results have been achieved with first generation direct-acting antivirals targeting the viral polymerase RdRp or the protease 3CL^pro . Such recently approved or investigational drugs like remdesivir and GC376 represent a basis for further development and optimization. Here, we establish a multi-readout assay (MRA) system that enables the antiviral assessment and mechanistic characterization of novel test compounds, drug repurposing and combination treatments. Our SARS-CoV-2-specific MRA combines the quantitative measurement of several parameters of virus infection, such as the intracellular production of proteins and genomes, enzymatic activities and virion release, as well as the use of reporter systems. In this regard, the antiviral efficacy of remdesivir and GC376 has been investigated in human Caco-2 cells. The readouts included the use of spike-and double-strand RNA-specific monoclonal antibodies for in-cell fluorescence imaging, a newly generated recombinant SARS-CoV-2 reporter virus d6YFP, the novel 3CL^pro-based FRET CFP::YFP and the previously reported FlipGFP reporter assays, as well as viral genome-specific RT-qPCR. The data produced by our MRA confirm the high antiviral potency of these two drugs in vitro. Combined, this MRA approach may be applied for broader analyses of SARS-CoV-2-specific antivirals, including compound screenings and the characterization of selected drug candidates.

Authors with CRIS profile

Friedrich Hahn Professur für Virologie Sigrun Häge Institute of Clinical and Molecular Virology Alexandra Herrmann Institute of Clinical and Molecular Virology Jintawee Kicuntod Professur für Pharmazeutische Chemie Klaus Überla Lehrstuhl für Klinische und Molekulare Virologie Armin Enßer Medizinische Fakultät Manfred Marschall Lehrstuhl für Klinische und Molekulare Virologie

Involved external institutions

Immunic AG

Germany (DE)

How to cite

APA:

Hahn, F., Häge, S., Herrmann, A., Wangen, C., Kicuntod, J., Jungnickl, D.,... Marschall, M. (2021). Methodological development of a multi-readout assay for the assessment of antiviral drugs against sars-cov-2. Pathogens, 10(9). https://dx.doi.org/10.3390/pathogens10091076

MLA:

Hahn, Friedrich, et al. "Methodological development of a multi-readout assay for the assessment of antiviral drugs against sars-cov-2." Pathogens 10.9 (2021).

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