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Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy
The efficacy of an antibody-drug conjugate (ADC) is dependent on the properties of its linker-payload which must remain stable while in systemic circulation but undergo efficient processing upon internalization into target cells. Here, we examine the stability of a non-cleavable Amino-PEG6-based lin...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498778/ https://www.ncbi.nlm.nih.gov/pubmed/26161543 http://dx.doi.org/10.1371/journal.pone.0132282 |
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author | Dorywalska, Magdalena Strop, Pavel Melton-Witt, Jody A. Hasa-Moreno, Adela Farias, Santiago E. Galindo Casas, Meritxell Delaria, Kathy Lui, Victor Poulsen, Kris Sutton, Janette Bolton, Gary Zhou, Dahui Moine, Ludivine Dushin, Russell Tran, Thomas-Toan Liu, Shu-Hui Rickert, Mathias Foletti, Davide Shelton, David L. Pons, Jaume Rajpal, Arvind |
author_facet | Dorywalska, Magdalena Strop, Pavel Melton-Witt, Jody A. Hasa-Moreno, Adela Farias, Santiago E. Galindo Casas, Meritxell Delaria, Kathy Lui, Victor Poulsen, Kris Sutton, Janette Bolton, Gary Zhou, Dahui Moine, Ludivine Dushin, Russell Tran, Thomas-Toan Liu, Shu-Hui Rickert, Mathias Foletti, Davide Shelton, David L. Pons, Jaume Rajpal, Arvind |
author_sort | Dorywalska, Magdalena |
collection | PubMed |
description | The efficacy of an antibody-drug conjugate (ADC) is dependent on the properties of its linker-payload which must remain stable while in systemic circulation but undergo efficient processing upon internalization into target cells. Here, we examine the stability of a non-cleavable Amino-PEG6-based linker bearing the monomethyl auristatin D (MMAD) payload site-specifically conjugated at multiple positions on an antibody. Enzymatic conjugation with transglutaminase allows us to create a stable amide linkage that remains intact across all tested conjugation sites on the antibody, and provides us with an opportunity to examine the stability of the auristatin payload itself. We report a position-dependent degradation of the C terminus of MMAD in rodent plasma that has a detrimental effect on its potency. The MMAD cleavage can be eliminated by either modifying the C terminus of the toxin, or by selection of conjugation site. Both approaches result in improved stability and potency in vitro and in vivo. Furthermore, we show that the MMAD metabolism in mouse plasma is likely mediated by a serine-based hydrolase, appears much less pronounced in rat, and was not detected in cynomolgus monkey or human plasma. Clarifying these species differences and controlling toxin degradation to optimize ADC stability in rodents is essential to make the best ADC selection from preclinical models. The data presented here demonstrate that site selection and toxin susceptibility to mouse plasma degradation are important considerations in the design of non-cleavable ADCs, and further highlight the benefits of site-specific conjugation methods. |
format | Online Article Text |
id | pubmed-4498778 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-44987782015-07-17 Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy Dorywalska, Magdalena Strop, Pavel Melton-Witt, Jody A. Hasa-Moreno, Adela Farias, Santiago E. Galindo Casas, Meritxell Delaria, Kathy Lui, Victor Poulsen, Kris Sutton, Janette Bolton, Gary Zhou, Dahui Moine, Ludivine Dushin, Russell Tran, Thomas-Toan Liu, Shu-Hui Rickert, Mathias Foletti, Davide Shelton, David L. Pons, Jaume Rajpal, Arvind PLoS One Research Article The efficacy of an antibody-drug conjugate (ADC) is dependent on the properties of its linker-payload which must remain stable while in systemic circulation but undergo efficient processing upon internalization into target cells. Here, we examine the stability of a non-cleavable Amino-PEG6-based linker bearing the monomethyl auristatin D (MMAD) payload site-specifically conjugated at multiple positions on an antibody. Enzymatic conjugation with transglutaminase allows us to create a stable amide linkage that remains intact across all tested conjugation sites on the antibody, and provides us with an opportunity to examine the stability of the auristatin payload itself. We report a position-dependent degradation of the C terminus of MMAD in rodent plasma that has a detrimental effect on its potency. The MMAD cleavage can be eliminated by either modifying the C terminus of the toxin, or by selection of conjugation site. Both approaches result in improved stability and potency in vitro and in vivo. Furthermore, we show that the MMAD metabolism in mouse plasma is likely mediated by a serine-based hydrolase, appears much less pronounced in rat, and was not detected in cynomolgus monkey or human plasma. Clarifying these species differences and controlling toxin degradation to optimize ADC stability in rodents is essential to make the best ADC selection from preclinical models. The data presented here demonstrate that site selection and toxin susceptibility to mouse plasma degradation are important considerations in the design of non-cleavable ADCs, and further highlight the benefits of site-specific conjugation methods. Public Library of Science 2015-07-10 /pmc/articles/PMC4498778/ /pubmed/26161543 http://dx.doi.org/10.1371/journal.pone.0132282 Text en © 2015 Dorywalska et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Dorywalska, Magdalena Strop, Pavel Melton-Witt, Jody A. Hasa-Moreno, Adela Farias, Santiago E. Galindo Casas, Meritxell Delaria, Kathy Lui, Victor Poulsen, Kris Sutton, Janette Bolton, Gary Zhou, Dahui Moine, Ludivine Dushin, Russell Tran, Thomas-Toan Liu, Shu-Hui Rickert, Mathias Foletti, Davide Shelton, David L. Pons, Jaume Rajpal, Arvind Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy |
title | Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy |
title_full | Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy |
title_fullStr | Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy |
title_full_unstemmed | Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy |
title_short | Site-Dependent Degradation of a Non-Cleavable Auristatin-Based Linker-Payload in Rodent Plasma and Its Effect on ADC Efficacy |
title_sort | site-dependent degradation of a non-cleavable auristatin-based linker-payload in rodent plasma and its effect on adc efficacy |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498778/ https://www.ncbi.nlm.nih.gov/pubmed/26161543 http://dx.doi.org/10.1371/journal.pone.0132282 |
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