Inhibitors of dihydroorotate dehydrogenase cooperate with molnupiravir and N4-hydroxycytidine to suppress SARS-CoV-2 replication
Kim M. Stegmann,
Antje Dickmanns,
Natalie Heinen,
Claudia Blaurock,
Tim Karrasch,
Angele Breithaupt,
Robert Klopfleisch,
Nadja Uhlig,
Valentina Eberlein,
Leila Issmail,
Simon T. Herrmann,
Amelie Schreieck,
Evelyn Peelen,
Hella Kohlhof,
Balal Sadeghi,
Alexander Riek,
John R. Speakman,
Uwe Groß,
Dirk Görlich,
Daniel Vitt,
Thorsten Müller,
Thomas Grunwald,
Stephanie Pfaender,
Anne Balkema-Buschmann,
Matthias Dobbelstein
Affiliations
Kim M. Stegmann
Institute of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, Justus von Liebig Weg 11, 37077 Göttingen, Germany
Antje Dickmanns
Institute of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, Justus von Liebig Weg 11, 37077 Göttingen, Germany
Natalie Heinen
Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
Claudia Blaurock
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany
Tim Karrasch
Institute of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, Justus von Liebig Weg 11, 37077 Göttingen, Germany
Angele Breithaupt
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany
Robert Klopfleisch
Institute of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany
Nadja Uhlig
Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany
Valentina Eberlein
Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany
Leila Issmail
Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany
Simon T. Herrmann
Department of Molecular Biochemistry, Ruhr University Bochum, Bochum, Germany
Amelie Schreieck
Immunic AG, Gräfelfing, Germany
Evelyn Peelen
Immunic AG, Gräfelfing, Germany
Hella Kohlhof
Immunic AG, Gräfelfing, Germany
Balal Sadeghi
Friedrich-Loeffler-Institut, Institute of Novel and Emerging Infectious Diseases, Greifswald-Insel Riems, Germany
Alexander Riek
Friedrich-Loeffler-Institut, Institute of Animal Welfare and Animal Husbandry, Celle, Germany
John R. Speakman
Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen, UK
Uwe Groß
Institute of Medical Microbiology and Virology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, Göttingen, Germany
Dirk Görlich
Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
Daniel Vitt
Immunic AG, Gräfelfing, Germany
Thorsten Müller
Department of Molecular Biochemistry, Ruhr University Bochum, Bochum, Germany; Institute of Psychiatric Phenomics and Genomics (IPPG), Organoid Laboratory, University Hospital, LMU Munich, Munich, Germany
Thomas Grunwald
Fraunhofer Institute for Cell Therapy and Immunology IZI, Leipzig, Germany
Stephanie Pfaender
Department of Molecular and Medical Virology, Ruhr University Bochum, Bochum, Germany
Anne Balkema-Buschmann
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany
Matthias Dobbelstein
Institute of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, Justus von Liebig Weg 11, 37077 Göttingen, Germany; Corresponding author
Summary: The nucleoside analog N4-hydroxycytidine (NHC) is the active metabolite of the prodrug molnupiravir, which has been approved for the treatment of COVID-19. SARS-CoV-2 incorporates NHC into its RNA, resulting in defective virus genomes. Likewise, inhibitors of dihydroorotate dehydrogenase (DHODH) reduce virus yield upon infection, by suppressing the cellular synthesis of pyrimidines. Here, we show that NHC and DHODH inhibitors strongly synergize in the inhibition of SARS-CoV-2 replication in vitro. We propose that the lack of available pyrimidine nucleotides upon DHODH inhibition increases the incorporation of NHC into nascent viral RNA. This concept is supported by the rescue of virus replication upon addition of pyrimidine nucleosides to the media. DHODH inhibitors increased the antiviral efficiency of molnupiravir not only in organoids of human lung, but also in Syrian Gold hamsters and in K18-hACE2 mice. Combining molnupiravir with DHODH inhibitors may thus improve available therapy options for COVID-19.
