Potent Anti-SARS-CoV-2 Activity by the Natural Product Gallinamide A and Analogues via Inhibition of Cathepsin L

Anneliese S Ashhurst; Arthur H Tang; Pavla Fajtová; Michael C Yoon; Anupriya Aggarwal; Max J Bedding; Alexander Stoye; Laura Beretta; Dustin Pwee; Aleksandra Drelich; Danielle Skinner; Linfeng Li; Thomas D Meek; James H McKerrow; Vivian Hook; Chien-Te Tseng; Mark Larance; Stuart Turville; William H Gerwick; Anthony J O'Donoghue; Richard J Payne

doi:10.1021/acs.jmedchem.1c01494

Potent Anti-SARS-CoV-2 Activity by the Natural Product Gallinamide A and Analogues via Inhibition of Cathepsin L

J Med Chem. 2022 Feb 24;65(4):2956-2970. doi: 10.1021/acs.jmedchem.1c01494. Epub 2021 Nov 3.

Authors

Anneliese S Ashhurst^{1

2}, Arthur H Tang¹, Pavla Fajtová^{3

4}, Michael C Yoon³, Anupriya Aggarwal⁵, Max J Bedding¹, Alexander Stoye¹, Laura Beretta³, Dustin Pwee³, Aleksandra Drelich⁶, Danielle Skinner³, Linfeng Li⁷, Thomas D Meek⁷, James H McKerrow³, Vivian Hook³, Chien-Te Tseng⁶, Mark Larance⁸, Stuart Turville⁵, William H Gerwick^{3

9}, Anthony J O'Donoghue³, Richard J Payne^{1

10}

Affiliations

¹ School of Chemistry, The University of Sydney, Sydney, NSW2006, Australia.
² School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW2006, Australia.
³ Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, California92093, United States.
⁴ Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, 16610Prague, Czech Republic.
⁵ Kirby Institute, University of New South Wales, Sydney, NSW2052, Australia.
⁶ Department of Microbiology and Immunology, University of Texas, Medical Branch, 3000 University Boulevard, Galveston, Texas77755-1001, United States.
⁷ Department of Biochemistry and Biophysics, Texas A&M University, 301 Old Main Drive, College Station, Texas77843, United States.
⁸ Charles Perkins Centre and School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW2006, Australia.
⁹ Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, California92093, United States.
¹⁰ Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Sydney, Sydney, NSW2006, Australia.

Abstract

Cathepsin L is a key host cysteine protease utilized by coronaviruses for cell entry and is a promising drug target for novel antivirals against SARS-CoV-2. The marine natural product gallinamide A and several synthetic analogues were identified as potent inhibitors of cathepsin L with IC₅₀ values in the picomolar range. Lead molecules possessed selectivity over other cathepsins and alternative host proteases involved in viral entry. Gallinamide A directly interacted with cathepsin L in cells and, together with two lead analogues, potently inhibited SARS-CoV-2 infection in vitro, with EC₅₀ values in the nanomolar range. Reduced antiviral activity was observed in cells overexpressing transmembrane protease, serine 2 (TMPRSS2); however, a synergistic improvement in antiviral activity was achieved when combined with a TMPRSS2 inhibitor. These data highlight the potential of cathepsin L as a COVID-19 drug target as well as the likely need to inhibit multiple routes of viral entry to achieve efficacy.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

A549 Cells
Animals
Antimicrobial Cationic Peptides / chemical synthesis
Antimicrobial Cationic Peptides / chemistry
Antimicrobial Cationic Peptides / pharmacology*
Antiviral Agents / chemical synthesis
Antiviral Agents / chemistry
Antiviral Agents / pharmacology*
Biological Products / chemical synthesis
Biological Products / chemistry
Biological Products / pharmacology*
COVID-19 / metabolism
COVID-19 Drug Treatment*
Cathepsin L / antagonists & inhibitors*
Cathepsin L / metabolism
Chlorocebus aethiops
Cysteine Proteinase Inhibitors / chemical synthesis
Cysteine Proteinase Inhibitors / chemistry
Cysteine Proteinase Inhibitors / pharmacology*
Dose-Response Relationship, Drug
Humans
Microbial Sensitivity Tests
Molecular Conformation
Proteomics
SARS-CoV-2 / drug effects*
Structure-Activity Relationship
Vero Cells

Substances

Antimicrobial Cationic Peptides
Antiviral Agents
Biological Products
Cysteine Proteinase Inhibitors
gallinamide A
CTSL protein, human
Cathepsin L

Grants and funding

U01 TW006634/TW/FIC NIH HHS/United States