Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
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Analysis of SARS-CoV-2 Mutations after Nirmatrelvir Treatment in a Lung Cancer Xenograft Mouse Model

  • Bo Min Kang (Department of Microbiology, College of Medicine, Hallym University) ;
  • Dongbum Kim (Institute of Medical Science, College of Medicine, Hallym University) ;
  • Jinsoo Kim (Institute of Medical Science, College of Medicine, Hallym University) ;
  • Kyeongbin Baek (Department of Microbiology, College of Medicine, Hallym University) ;
  • Sangkyu Park (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Ha-Eun Shin (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Myeong-Heon Lee (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Minyoung Kim (Department of Microbiology, College of Medicine, Hallym University) ;
  • Suyeon Kim (Department of Microbiology, College of Medicine, Hallym University) ;
  • Younghee Lee (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Hyung-Joo Kwon (Department of Microbiology, College of Medicine, Hallym University)
  • Received : 2023.11.06
  • Accepted : 2023.12.18
  • Published : 2024.07.01
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Abstract

Paxlovid is the first approved oral treatment for coronavirus disease 2019 and includes nirmatrelvir, a protease inhibitor targeting the main protease (Mpro) of SARS-CoV-2, as one of the key components. While some specific mutations emerged in Mpro were revealed to significantly reduce viral susceptibility to nirmatrelvir in vitro, there is no report regarding resistance to nirmatrelvir in patients and animal models for SARS-CoV-2 infection yet. We recently developed xenograft tumors derived from Calu-3 cells in immunodeficient mice and demonstrated extended replication of SARS-CoV-2 in the tumors. In this study, we investigated the effect of nirmatrelvir administration on SARS-CoV-2 replication. Treatment with nirmatrelvir after virus infection significantly reduced the replication of the parental SARS-CoV-2 and SARS-CoV-2 Omicron at 5 days post-infection (dpi). However, the virus titers were completely recovered at the time points of 15 and 30 dpi. The virus genomes in the tumors at 30 dpi were analyzed to investigate whether nirmatrelvir-resistant mutant viruses had emerged during the extended replication of SARS-CoV-2. Various mutations in several genes including ORF1ab, ORF3a, ORF7a, ORF7b, ORF8, and N occurred in the SARS-CoV-2 genome; however, no mutations were induced in the Mpro sequence by a single round of nirmatrelvir treatment, and none were observed even after two rounds of treatment. The parental SARS-CoV-2 and its sublineage isolates showed similar IC50 values of nirmatrelvir in Vero E6 cells. Therefore, it is probable that inducing viral resistance to nirmatrelvir in vivo is challenging differently from in vitro passage.

Keywords

Acknowledgement

We thank the National Culture Collection for Pathogens (Osong, Korea) for supplying parental SARS-CoV-2 and SARS-CoV-2 Omicron. This research was supported by grants from the National Research Foundation (NRF-2022M3A9I2082292) funded by the Ministry of Science and ICT in the Republic of Korea and by a grant of the Korea Health Technology R&D Project (grant number: HV23C0052) through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare in the Republic of Korea.

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