Diphyllin elicits a doubled-pronged attack on the entry of SARS-CoV-2 by inhibiting cathepsin L and furin

  • Virus Res. 2024 Oct 19:350:199485. doi: 10.1016/j.virusres.2024.199485.
Binli Mao  1 Vu Thuy Khanh Le-Trilling  2 Haihuan Tang  3 Jie Hu  4 Mona S Schmitz  5 Kimberly Barbet  5 Dan Xu  3 Zhen Wei  3 Beinu Guo  3 Denise Mennerich  6 Chun Yao  7 Jinxin Liu  7 Zhenghan Li  7 Yushun Wan  8 Xiaoyong Zhang  9 Kai Wang  3 Ni Tang  3 Zebo Yu  10 Mirko Trilling  11 Yong Lin  12
Affiliations
  • 1. Key Laboratory of Molecular Biology of Infectious Diseases (Chinese Ministry of Education), Chongqing Medical University, Chongqing 400016, China; Department of Blood Transfusion, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
  • 2. Institute for the Research on HIV and AIDS-associated Diseases, University Hospital Essen, University of Duisburg-Essen, Essen 45147, Germany; Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen 45147, Germany.
  • 3. Key Laboratory of Molecular Biology of Infectious Diseases (Chinese Ministry of Education), Chongqing Medical University, Chongqing 400016, China.
  • 4. Key Laboratory of Molecular Biology of Infectious Diseases (Chinese Ministry of Education), Chongqing Medical University, Chongqing 400016, China; Department of Laboratory Medicine, Bishan Hospital of Chongqing Medical University, Chongqing 402760, China.
  • 5. Department of Pulmonary Medicine, University Medical Center Essen, Ruhrlandklinik, Essen 45239, Germany.
  • 6. Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen 45147, Germany.
  • 7. Chongqing Yucai Secondary School, Chongqing 400050, China.
  • 8. College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
  • 9. Hepatology Unit and Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
  • 10. Department of Blood Transfusion, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China. Electronic address: [email protected].
  • 11. Institute for the Research on HIV and AIDS-associated Diseases, University Hospital Essen, University of Duisburg-Essen, Essen 45147, Germany; Institute for Virology, University Hospital Essen, University of Duisburg-Essen, Essen 45147, Germany. Electronic address: [email protected].
  • 12. Key Laboratory of Molecular Biology of Infectious Diseases (Chinese Ministry of Education), Chongqing Medical University, Chongqing 400016, China. Electronic address: [email protected].
Abstract

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the coronavirus disease 2019 (COVID-19) pandemic, posing serious threats to global health. Effective broad-spectrum Antiviral drugs for the treatment of COVID-19 are not sufficiently available. In the present study, we investigated the Antiviral activity of the natural lignan diphyllin (PubChem CID 100492) against different SARS-CoV-2 variants and explored the underlying molecular mechanisms. We found that diphyllin dose-dependently inhibits the SARS-CoV-2 spike (S)-mediated entry into different types of cells. The potent inhibition was evident against spike proteins derived from the original SARS-CoV-2 and from variants of concern such as Alpha, Beta, Delta or Omicron. Accordingly, diphyllin also significantly inhibited the in vitro Infection of a clinical SARS-CoV-2 virus isolate. Mechanistically, diphyllin simultaneously inhibited the endosomal entry of SARS-CoV-2 by neutralizing the endosomal acidification and reducing the activity of the cysteine protease Cathepsin L (CTSL) as well as S-meditated cell surface entry by impairing Furin activity. Collectively, our findings establish diphyllin as novel inhibitor of CTSL and Furin proteases, resulting in a double-pronged attack on SARS-CoV-2 entry along endosomal as well as cell surface routes. Therefore, diphyllin has the potential to be advanced as an inhibitor of SARS-CoV-2 entry.

Keywords
Cathepsin L; Cell surface entry; Diphyllin; Endocytosis; Furin; SARS-CoV-2.
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