Exposure of human intestinal epithelial cells and primary human hepatocytes to trypsin-like serine protease inhibitors with potential antiviral effect

J Enzyme Inhib Med Chem. 2021 Dec;36(1):659-668. doi: 10.1080/14756366.2021.1886093.

Abstract

Human intestinal epithelial cell line-6 (HIEC-6) cells and primary human hepatocytes (PHHs) were treated with 3-amidinophenylalanine-derived inhibitors of trypsin-like serine proteases for 24 hours. It was proven that treatment with MI-1900 and MI-1907 was tolerated up to 50 μM in HIEC-6. These inhibitors did not cause elevations in extracellular H2O2 levels and in the concentrations of interleukin (IL)-6 and IL-8 and did not alter occludin distribution in HIEC-6. It was also found that MI-1900 and MI-1907 up to 50 μM did not affect cell viability, IL-6 and IL-8 and occludin levels of PHH. Based on our findings, these inhibitors could be safely applicable at 50 μM in HIEC-6 and in PHH; however, redox status was disturbed in case of PHH. Moreover, it has recently been demonstrated that MI-1900 prevents the replication and spread of the new SARS-CoV-2 in infected Calu-3 cells, most-likely via an inhibition of the membrane-bound host protease TMPRSS2.

Keywords: H2O2; HIEC-6; PHH; occludin; trypsin-like serine protease inhibitors.

MeSH terms

  • Antiviral Agents / pharmacology*
  • Cell Line
  • Cell Survival / drug effects
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects*
  • Epithelial Cells / enzymology
  • Gene Expression Regulation / drug effects
  • Hepatocytes / cytology
  • Hepatocytes / drug effects*
  • Hepatocytes / enzymology
  • Humans
  • Hydrogen Peroxide / metabolism
  • Interleukin-6 / genetics
  • Interleukin-6 / metabolism
  • Interleukin-8 / genetics
  • Interleukin-8 / metabolism
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / enzymology
  • Occludin / genetics
  • Occludin / metabolism
  • Oxidation-Reduction / drug effects
  • Phenylalanine / analogs & derivatives
  • Phenylalanine / pharmacology*
  • Primary Cell Culture
  • Protease Inhibitors / pharmacology*
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism*

Substances

  • Antiviral Agents
  • IL6 protein, human
  • Interleukin-6
  • Interleukin-8
  • OCLN protein, human
  • Occludin
  • Protease Inhibitors
  • trypsin-like serine protease
  • Phenylalanine
  • Hydrogen Peroxide
  • Serine Endopeptidases
  • TMPRSS2 protein, human

Grants and funding

This work was supported by the Hungarian National Research, Development and Innovation Office under Grant 115685 and 124522; and the European Union and Co-Financed by the European Social Fund under Grant Agreement No. EFOP-3.6.1-16-2016-00024 and EFOP-3.6.3-VEKOP-16-2017-00005, project title: Strengthening the scientific replacement by supporting the academic workshops and programs of students, developing a mentoring process. This project was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences and by the ÚNKP-20-5 New National Excellence Program of the Ministry for Innovation and Technology from the Source of the National Research, Development and Innovation Fund.