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In vitro selection of macrocyclic peptide inhibitors containing cyclic γ2,4-amino acids targeting the SARS-CoV-2 main protease

Abstract:
γ-Amino acids can play important roles in the biological activities of natural products; however, ribosomal incorporation of γ-amino acids into peptides is challenging. Here we report how a selection campaign employing a non-canonical peptide library containing cyclic γ2,4-amino acids (cγAAs) resulted in the discovery of very potent inhibitors of the SARS-CoV-2 main protease (Mpro). Two kinds of cγAAs, cis-3-aminocyclobutane carboxylic acid (γ1) and (1R,3S)-3-aminocyclopentane carboxylic acid (γ2), were ribosomally introduced into a library of thioether-macrocyclic peptides. One resultant potent Mpro inhibitor (IC50 = 50 nM), GM4, comprising 13 residues with γ1 at the 4th position, manifests a 5.2 nM dissociation constant. An Mpro:GM4 complex crystal structure reveals the intact inhibitor spans the substrate binding cleft. γ1 interacts with the S1′ catalytic subsite and contributes to a 12-fold increase in proteolytic stability compared to its alanine-substituted variant. Knowledge of interactions between GM4 and Mpro enabled production of a variant with a 5-fold increase in potency.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/s41557-023-01205-1

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Publisher:
Springer Nature
Journal:
Nature Chemistry More from this journal
Volume:
15
Pages:
998–1005
Publication date:
2023-05-22
Acceptance date:
2023-04-14
DOI:
EISSN:
1755-4349
ISSN:
1755-4330


Language:
English
Keywords:
Pubs id:
1331038
Local pid:
pubs:1331038
Deposit date:
2023-03-02
ARK identifier:

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