Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp5 main protease.
Amino Acid Chloromethyl Ketones
/ pharmacology
Animals
Antiviral Agents
/ chemistry
Azoles
/ pharmacology
Chlorocebus aethiops
Coronavirus 3C Proteases
/ antagonists & inhibitors
Drug Evaluation, Preclinical
Enzyme Assays
Fluorescence Resonance Energy Transfer
High-Throughput Screening Assays
Isoindoles
Leupeptins
/ pharmacology
Organoselenium Compounds
/ pharmacology
Peptidomimetics
RNA-Binding Proteins
/ metabolism
Reproducibility of Results
SARS-CoV-2
/ drug effects
Small Molecule Libraries
/ chemistry
Vero Cells
Viral Nonstructural Proteins
/ metabolism
COVID-19
coronavirus
nsp5
protease
Journal
The Biochemical journal
ISSN: 1470-8728
Titre abrégé: Biochem J
Pays: England
ID NLM: 2984726R
Informations de publication
Date de publication:
16 07 2021
16 07 2021
Historique:
received:
23
03
2021
revised:
05
05
2021
accepted:
07
05
2021
entrez:
1
7
2021
pubmed:
2
7
2021
medline:
9
7
2021
Statut:
ppublish
Résumé
The coronavirus 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spread around the world with unprecedented health and socio-economic effects for the global population. While different vaccines are now being made available, very few antiviral drugs have been approved. The main viral protease (nsp5) of SARS-CoV-2 provides an excellent target for antivirals, due to its essential and conserved function in the viral replication cycle. We have expressed, purified and developed assays for nsp5 protease activity. We screened the nsp5 protease against a custom chemical library of over 5000 characterised pharmaceuticals. We identified calpain inhibitor I and three different peptidyl fluoromethylketones (FMK) as inhibitors of nsp5 activity in vitro, with IC50 values in the low micromolar range. By altering the sequence of our peptidomimetic FMK inhibitors to better mimic the substrate sequence of nsp5, we generated an inhibitor with a subnanomolar IC50. Calpain inhibitor I inhibited viral infection in monkey-derived Vero E6 cells, with an EC50 in the low micromolar range. The most potent and commercially available peptidyl-FMK compound inhibited viral growth in Vero E6 cells to some extent, while our custom peptidyl FMK inhibitor offered a marked antiviral improvement.
Identifiants
pubmed: 34198327
pii: 229152
doi: 10.1042/BCJ20210197
pmc: PMC8286836
doi:
Substances chimiques
Amino Acid Chloromethyl Ketones
0
Antiviral Agents
0
Azoles
0
Isoindoles
0
Leupeptins
0
NSP9 protein, SARS-CoV-2
0
Organoselenium Compounds
0
Peptidomimetics
0
RNA-Binding Proteins
0
Small Molecule Libraries
0
Viral Nonstructural Proteins
0
acetylleucyl-leucyl-norleucinal
110044-82-1
ebselen
40X2P7DPGH
3C-like proteinase, SARS-CoV-2
EC 3.4.22.-
Coronavirus 3C Proteases
EC 3.4.22.28
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2499-2515Subventions
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Cancer Research UK
ID : 24558
Pays : United Kingdom
Organisme : Arthritis Research UK
ID : FC001066
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UU_00018/4
Pays : United Kingdom
Informations de copyright
© 2021 The Author(s).
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