Photochemically-enabled, post-translational production of C-terminal amides

Abstract C-terminal α-amidated peptides are attractive therapeutic targets, but preparative methods to access amidated pharmaceuticals are limited both on lab and manufacturing-scale. Here we report a straightforward and scalable approach to the C-terminal α-amidation of peptides and proteins from c...

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Main Authors: David Hymel, Felix Wojcik, Kim S. Halskov, Wouter F. J. Hogendorf, Sydnee C. Wong, Ben M. Williams, Asmus R. Mortensen, Nick Cox, Ayesha Misquith, Nanna B. Holländer, Finn Matthiesen, Suneet Mehrotra, Michael R. Harris
Format: Article
Language:English
Published: Nature Portfolio 2024-11-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-024-51005-5
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author David Hymel
Felix Wojcik
Kim S. Halskov
Wouter F. J. Hogendorf
Sydnee C. Wong
Ben M. Williams
Asmus R. Mortensen
Nick Cox
Ayesha Misquith
Nanna B. Holländer
Finn Matthiesen
Suneet Mehrotra
Michael R. Harris
author_facet David Hymel
Felix Wojcik
Kim S. Halskov
Wouter F. J. Hogendorf
Sydnee C. Wong
Ben M. Williams
Asmus R. Mortensen
Nick Cox
Ayesha Misquith
Nanna B. Holländer
Finn Matthiesen
Suneet Mehrotra
Michael R. Harris
author_sort David Hymel
collection DOAJ
description Abstract C-terminal α-amidated peptides are attractive therapeutic targets, but preparative methods to access amidated pharmaceuticals are limited both on lab and manufacturing-scale. Here we report a straightforward and scalable approach to the C-terminal α-amidation of peptides and proteins from cysteine-extended polypeptide precursors. This amidation protocol consists of three highly efficient steps: 1) selective cysteine thiol substitution with a photolabel, 2) photoinduced decarboxylative elimination and 3) enamide cleavage by simple acidolysis or inverse electron demand Diels-Alder reaction. We provide a blueprint for applying this protocol to the semi-recombinant production of therapeutically relevant targets where gram scale C-terminal α-amidation is achieved in a photoflow reactor on a recombinantly prepared peptide YY analogue and a GLP-1/amylin co-agonist precursor peptide. Robust performance of this reaction cascade in flow highlights the potential of this chemistry to enable amidated drug leads to enter development that would not be viable on commercial scale using existing technology.
format Article
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institution Kabale University
issn 2041-1723
language English
publishDate 2024-11-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-9979539e39a44b69b33db07562ed58032024-12-01T12:34:52ZengNature PortfolioNature Communications2041-17232024-11-011511810.1038/s41467-024-51005-5Photochemically-enabled, post-translational production of C-terminal amidesDavid Hymel0Felix Wojcik1Kim S. Halskov2Wouter F. J. Hogendorf3Sydnee C. Wong4Ben M. Williams5Asmus R. Mortensen6Nick Cox7Ayesha Misquith8Nanna B. Holländer9Finn Matthiesen10Suneet Mehrotra11Michael R. Harris12Novo Nordisk Research Center Seattle, Inc.Research Chemistry, Novo Nordisk A/SResearch Chemistry, Novo Nordisk A/SResearch Chemistry, Novo Nordisk A/SNovo Nordisk Research Center Seattle, Inc.Chemical Development, Novo Nordisk A/SChemical Development, Novo Nordisk A/SNovo Nordisk Research Center Seattle, Inc.Novo Nordisk Research Center Seattle, Inc.Chemical Development, Novo Nordisk A/SPurification Technologies, Novo Nordisk A/SNovo Nordisk Research Center Seattle, Inc.Novo Nordisk Research Center Seattle, Inc.Abstract C-terminal α-amidated peptides are attractive therapeutic targets, but preparative methods to access amidated pharmaceuticals are limited both on lab and manufacturing-scale. Here we report a straightforward and scalable approach to the C-terminal α-amidation of peptides and proteins from cysteine-extended polypeptide precursors. This amidation protocol consists of three highly efficient steps: 1) selective cysteine thiol substitution with a photolabel, 2) photoinduced decarboxylative elimination and 3) enamide cleavage by simple acidolysis or inverse electron demand Diels-Alder reaction. We provide a blueprint for applying this protocol to the semi-recombinant production of therapeutically relevant targets where gram scale C-terminal α-amidation is achieved in a photoflow reactor on a recombinantly prepared peptide YY analogue and a GLP-1/amylin co-agonist precursor peptide. Robust performance of this reaction cascade in flow highlights the potential of this chemistry to enable amidated drug leads to enter development that would not be viable on commercial scale using existing technology.https://doi.org/10.1038/s41467-024-51005-5
spellingShingle David Hymel
Felix Wojcik
Kim S. Halskov
Wouter F. J. Hogendorf
Sydnee C. Wong
Ben M. Williams
Asmus R. Mortensen
Nick Cox
Ayesha Misquith
Nanna B. Holländer
Finn Matthiesen
Suneet Mehrotra
Michael R. Harris
Photochemically-enabled, post-translational production of C-terminal amides
Nature Communications
title Photochemically-enabled, post-translational production of C-terminal amides
title_full Photochemically-enabled, post-translational production of C-terminal amides
title_fullStr Photochemically-enabled, post-translational production of C-terminal amides
title_full_unstemmed Photochemically-enabled, post-translational production of C-terminal amides
title_short Photochemically-enabled, post-translational production of C-terminal amides
title_sort photochemically enabled post translational production of c terminal amides
url https://doi.org/10.1038/s41467-024-51005-5
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