Reactive oxygen species-mediated cytotoxic and DNA-damaging mechanism of N4-hydroxycytidine, a metabolite of the COVID-19 therapeutic drug molnupiravir

Mori et al., Free Radical Research, doi:10.1080/10715762.2025.2469738, Feb 2025
In vitro study showing that molnupiravir may have cytotoxic and mutagenic effects in host cells via hydroxylamine production from N4-hydroxycytidine (NHC) by cytidine deaminase (CDA). Molnupiravir metabolite NHC may induce cytotoxicity and mutagenicity through CDA-mediated reactive oxygen species generation.
Potential risks of molnupiravir include the creation of dangerous variants, and mutagenicity, carcinogenicity, teratogenicity, and embryotoxicity1-15. Multiple analyses have identified variants potentially created by molnupiravir16-20. Studies show significantly increased risk of acute kidney injury21, cardiovascular toxocity22, and neurological symptoms21. Treatment may increase viral rebound23,24.
Mori et al., 20 Feb 2025, Japan, peer-reviewed, 10 authors. Contact: s-oikawa@med.mie-u.ac.jp.
In vitro studies are an important part of preclinical research, however results may be very different in vivo.
Abstract: Free Radical Research ISSN: (Print) (Online) Journal homepage: www.tandfonline.com/journals/ifra20 Reactive oxygen species-mediated cytotoxic and DNA-damaging mechanism of N4-hydroxycytidine, a metabolite of the COVID-19 therapeutic drug molnupiravir Yurie Mori, Rinya Yogo, Hatasu Kobayashi, Hirotaka Katsuzaki, Yuichiro Hirao, Shinya Kato, Hirokazu Kotani, Shosuke Kawanishi, Mariko Murata & Shinji Oikawa To cite this article: Yurie Mori, Rinya Yogo, Hatasu Kobayashi, Hirotaka Katsuzaki, Yuichiro Hirao, Shinya Kato, Hirokazu Kotani, Shosuke Kawanishi, Mariko Murata & Shinji Oikawa (20 4 Feb 2025): Reactive oxygen species-mediated cytotoxic and DNA-damaging mechanism of N hydroxycytidine, a metabolite of the COVID-19 therapeutic drug molnupiravir, Free Radical Research, DOI: 10.1080/10715762.2025.2469738 To link to this article: https://doi.org/10.1080/10715762.2025.2469738 © 2025 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group Accepted author version posted online: 20 Feb 2025. Submit your article to this journal View related articles View Crossmark data Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=ifra20 Revised manuscript Reactive oxygen species-mediated cytotoxic and DNA-damaging mechanism of N4-hydroxycytidine, a metabolite of the COVID-19 ip t therapeutic drug molnupiravir Yurie Mori1,#, Rinya Yogo1,2,#, Hatasu Kobayashi1, Hirotaka Katsuzaki3, Yuichiro cr Hirao1, Shinya Kato4, Hirokazu Kotani 2, Shosuke Kawanishi5, Mariko Murata1, Shinji an Yurie Mori and Rinya Yogo should be considered joint first authors. M # us Oikawa1* 1. Department of Environmental and Molecular Medicine, Mie University Graduate ed School of Medicine, Edobashi 2-174, Tsu, Mie 514-8507, Japan pt 2. Department of Forensic Medicine and Sciences, Mie University Graduate School of ce Medicine, Edobashi 2-174, Tsu, Mie 514-8507, Japan 3. Department of Life Sciences, Graduate School of Bioresources, Mie University, Ac 1577 Kurimamachiya, Tsu, Mie 514-8507, Japan 4. Radioisotope Experimental Facility, Advanced Science Research Promotion Center, Mie University, Edobashi 2-174, Tsu, Mie 514-8507, Japan 5. Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, 3500-3, Minamitamagaki, Suzuka, Mie, 513-8670, Japan 1 * Corresponding author at: Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Edobashi 2-174, Tsu, Mie 514-8507, Japan. E-mail address: s-oikawa@med.mie-u.ac.jp (S. Oikawa) Running title: ROS generation mechanism of hydroxycytidine Keywords: molnupiravir, N4-hydroxycytidine, cytidine deaminase, reactive oxygen Ac ce pt ed M an us cr ip t species, cytotoxicity, DNA damage 2 Abstract Molnupiravir is a prodrug of the antiviral ribonucleoside analogue N4-hydroxycytidine (NHC), for use in treatment of coronavirus disease 2019 (COVID-19). However, it is generally considered that NHC-triphosphate is incorporated into the host genome to ip t induce mutations. In our previous preliminary report, we proposed oxidative DNA cr damage by NHC via cytidine deaminase (CDA)-mediated ROS formation. In the us present study, we investigated cell viability using the HL-60 human leukemia cell line an and its H2O2-resistant clone, HP100 cells. The survival rate was significantly reduced in HL-60 cells treated with NHC, but not in HP100 cells. LC-MS..
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