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N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates
Herein, the application of N‐hydroxysuccinimide‐modified phosphonamidate building blocks for the incorporation of cysteine‐selective ethynylphosphonamidates into lysine residues of proteins, followed by thiol addition with small molecules and proteins, is reported. It is demonstrated that the buildi...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7003776/ https://www.ncbi.nlm.nih.gov/pubmed/31661184 http://dx.doi.org/10.1002/cbic.201900587 |
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author | Kasper, Marc‐André Gerlach, Marcus Schneider, Anselm F. L. Groneberg, Christiane Ochtrop, Philipp Boldt, Stefanie Schumacher, Dominik Helma, Jonas Leonhardt, Heinrich Christmann, Mathias Hackenberger, Christian P. R. |
author_facet | Kasper, Marc‐André Gerlach, Marcus Schneider, Anselm F. L. Groneberg, Christiane Ochtrop, Philipp Boldt, Stefanie Schumacher, Dominik Helma, Jonas Leonhardt, Heinrich Christmann, Mathias Hackenberger, Christian P. R. |
author_sort | Kasper, Marc‐André |
collection | PubMed |
description | Herein, the application of N‐hydroxysuccinimide‐modified phosphonamidate building blocks for the incorporation of cysteine‐selective ethynylphosphonamidates into lysine residues of proteins, followed by thiol addition with small molecules and proteins, is reported. It is demonstrated that the building blocks significantly lower undesired homo‐crosslinking side products that can occur with commonly applied succinimidyl 4‐(N‐maleimidomethyl)cyclohexane‐1‐carboxylate (SMCC) under physiological pH. The previously demonstrated stability of the phosphonamidate moiety additionally solves the problem of premature maleimide hydrolysis, which can hamper the efficiency of subsequent thiol addition. Furthermore, a method to separate the phosphonamidate enantiomers to be able to synthesize protein conjugates in a defined configuration has been developed. Finally, the building blocks are applied to the construction of functional antibody–drug conjugates, analogously to FDA‐approved, SMCC‐linked Kadcyla, and to the synthesis of a functional antibody–protein conjugate. |
format | Online Article Text |
id | pubmed-7003776 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-70037762020-02-10 N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates Kasper, Marc‐André Gerlach, Marcus Schneider, Anselm F. L. Groneberg, Christiane Ochtrop, Philipp Boldt, Stefanie Schumacher, Dominik Helma, Jonas Leonhardt, Heinrich Christmann, Mathias Hackenberger, Christian P. R. Chembiochem Full Papers Herein, the application of N‐hydroxysuccinimide‐modified phosphonamidate building blocks for the incorporation of cysteine‐selective ethynylphosphonamidates into lysine residues of proteins, followed by thiol addition with small molecules and proteins, is reported. It is demonstrated that the building blocks significantly lower undesired homo‐crosslinking side products that can occur with commonly applied succinimidyl 4‐(N‐maleimidomethyl)cyclohexane‐1‐carboxylate (SMCC) under physiological pH. The previously demonstrated stability of the phosphonamidate moiety additionally solves the problem of premature maleimide hydrolysis, which can hamper the efficiency of subsequent thiol addition. Furthermore, a method to separate the phosphonamidate enantiomers to be able to synthesize protein conjugates in a defined configuration has been developed. Finally, the building blocks are applied to the construction of functional antibody–drug conjugates, analogously to FDA‐approved, SMCC‐linked Kadcyla, and to the synthesis of a functional antibody–protein conjugate. John Wiley and Sons Inc. 2020-01-07 2020-01-15 /pmc/articles/PMC7003776/ /pubmed/31661184 http://dx.doi.org/10.1002/cbic.201900587 Text en © 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Full Papers Kasper, Marc‐André Gerlach, Marcus Schneider, Anselm F. L. Groneberg, Christiane Ochtrop, Philipp Boldt, Stefanie Schumacher, Dominik Helma, Jonas Leonhardt, Heinrich Christmann, Mathias Hackenberger, Christian P. R. N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates |
title |
N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates |
title_full |
N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates |
title_fullStr |
N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates |
title_full_unstemmed |
N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates |
title_short |
N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates |
title_sort | n‐hydroxysuccinimide‐modified ethynylphosphonamidates enable the synthesis of configurationally defined protein conjugates |
topic | Full Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7003776/ https://www.ncbi.nlm.nih.gov/pubmed/31661184 http://dx.doi.org/10.1002/cbic.201900587 |
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