2. Academic Validation
  3. Enoxaparin augments alpha-1-antitrypsin inhibition of TMPRSS2, a promising drug combination against COVID-19

Enoxaparin augments alpha-1-antitrypsin inhibition of TMPRSS2, a promising drug combination against COVID-19

  • Sci Rep. 2022 Mar 25;12(1):5207. doi: 10.1038/s41598-022-09133-9.
Xiyuan Bai  # 1 2 3 4,  Ashley M Buckle  # 5,  Eszter K Vladar 6,  Edward N Janoff 7 8,  Reeti Khare 9,  Diane Ordway 10,  David Beckham 8,  Lorelenn B Fornis 11,  Abraham Majluf-Cruz 12,  Randolph V Fugit 13,  Brian M Freed 14,  Soohyun Kim 15 16,  Robert A Sandhaus 11,  Edward D Chan 17 18 19 20
Affiliations

Affiliations

  • 1 Department of Medicine, Rocky Mountain Regional Veterans Affairs Medical Center, Aurora, CO, USA. BaiX@NJHealth.org.
  • 2 Department of Academic Affairs and Medicine, National Jewish Health, Denver, CO, USA. BaiX@NJHealth.org.
  • 3 Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA. BaiX@NJHealth.org.
  • 4 National Jewish Health, D509, Neustadt Building, 1400 Jackson Street, Denver, CO, 80206, USA. BaiX@NJHealth.org.
  • 5 Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia.
  • 6 Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
  • 7 Department of Medicine, Rocky Mountain Regional Veterans Affairs Medical Center, Aurora, CO, USA.
  • 8 Division of Infectious Diseases, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
  • 9 Mycobacteriology Laboratory, Advance Diagnostics, National Jewish Health, Denver, CO, USA.
  • 10 Department of Microbiology, Immunlogy, and Pathology, Colorado State University, Fort Collins, CO, USA.
  • 11 Department of Academic Affairs and Medicine, National Jewish Health, Denver, CO, USA.
  • 12 Unidad de Investigacion Medica en Trombosis, Hemostasia y Aterogenesis, Instituto Mexicano del Seguro Social, Mexico City, Mexico.
  • 13 Department of Pharmacy, Rocky Mountain Regional Veterans Affairs Medical Center, Aurora, CO, USA.
  • 14 Department of Immunology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
  • 15 Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology, Konkuk University, Seoul, South Korea.
  • 16 College of Veterinary Medicine, Konkuk University, Seoul, South Korea.
  • 17 Department of Medicine, Rocky Mountain Regional Veterans Affairs Medical Center, Aurora, CO, USA. ChanE@NJHealth.org.
  • 18 Department of Academic Affairs and Medicine, National Jewish Health, Denver, CO, USA. ChanE@NJHealth.org.
  • 19 Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA. ChanE@NJHealth.org.
  • 20 National Jewish Health, D509, Neustadt Building, 1400 Jackson Street, Denver, CO, 80206, USA. ChanE@NJHealth.org.
  • # Contributed equally.
PMID: 35338216 DOI: 10.1038/s41598-022-09133-9
Abstract

The cell surface serine protease Transmembrane Protease 2 (TMPRSS2) is required to cleave the spike protein of SARS-CoV-2 for viral entry into cells. We determined whether negatively-charged heparin enhanced TMPRSS2 inhibition by alpha-1-antitrypsin (AAT). TMPRSS2 activity was determined in HEK293T cells overexpressing TMPRSS2. We quantified Infection of primary human airway epithelial cells (hAEc) with human coronavirus 229E (HCoV-229E) by immunostaining for the nucleocapsid protein and by the plaque assay. Detailed molecular modeling was undertaken with the heparin-TMPRSS2-AAT ternary complex. Enoxaparin enhanced AAT inhibition of both TMPRSS2 activity and Infection of hAEc with HCoV-229E. Underlying these findings, detailed molecular modeling revealed that: (i) the reactive center loop of AAT adopts an inhibitory-competent conformation compared with the crystal structure of TMPRSS2 bound to an exogenous (nafamostat) or endogenous (HAI-2) TMPRSS2 inhibitor and (ii) negatively-charged heparin bridges adjacent electropositive patches at the TMPRSS2-AAT interface, neutralizing otherwise repulsive forces. In conclusion, enoxaparin enhances AAT inhibition of both TMPRSS2 and coronavirus Infection. Such host-directed therapy is less likely to be affected by SARS-CoV-2 mutations. Furthermore, given the known anti-inflammatory activities of both AAT and heparin, this form of treatment may target both the virus and the excessive inflammatory consequences of severe COVID-19.

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