Single Mutation on Trastuzumab Modulates the Stability of Antibody-Drug Conjugates Built Using Acetal-Based Linkers and Thiol-Maleimide Chemistry.
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Authors
Ferhati, Xhenti
Cabeza-Cabrerizo, Mar
Compañón, Ismael
Akkapeddi, Padma
Salaverri, Noelia
Laserna, Víctor
Murray, Thomas V
Ravn, Peter
Publication Date
2022-03-30Journal Title
J Am Chem Soc
ISSN
0002-7863
Publisher
American Chemical Society (ACS)
Volume
144
Issue
12
Pages
5284-5294
Language
eng
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Ferhati, X., Jiménez-Moreno, E., Hoyt, E. A., Salluce, G., Cabeza-Cabrerizo, M., Navo, C. D., Compañón, I., et al. (2022). Single Mutation on Trastuzumab Modulates the Stability of Antibody-Drug Conjugates Built Using Acetal-Based Linkers and Thiol-Maleimide Chemistry.. J Am Chem Soc, 144 (12), 5284-5294. https://doi.org/10.1021/jacs.1c07675
Description
Funder: Universidad de La Rioja
Abstract
Antibody-drug conjugates (ADCs) are a class of targeted therapeutics used to selectively kill cancer cells. It is important that they remain intact in the bloodstream and release their payload in the target cancer cell for maximum efficacy and minimum toxicity. The development of effective ADCs requires the study of factors that can alter the stability of these therapeutics at the atomic level. Here, we present a general strategy that combines synthesis, bioconjugation, linker technology, site-directed mutagenesis, and modeling to investigate the influence of the site and microenvironment of the trastuzumab antibody on the stability of the conjugation and linkers. Trastuzumab is widely used to produce targeted ADCs because it can target with high specificity a receptor that is overexpressed in certain breast cancer cells (HER2). We show that the chemical environment of the conjugation site of trastuzumab plays a key role in the stability of linkers featuring acid-sensitive groups such as acetals. More specifically, Lys-207, located near the reactive Cys-205 of a thiomab variant of the antibody, may act as an acid catalyst and promote the hydrolysis of acetals. Mutation of Lys-207 into an alanine or using a longer linker that separates this residue from the acetal group stabilizes the conjugates. Analogously, Lys-207 promotes the beneficial hydrolysis of the succinimide ring when maleimide reagents are used for conjugation, thus stabilizing the subsequent ADCs by impairing the undesired retro-Michael reactions. This work provides new insights for the design of novel ADCs with improved stability properties.
Keywords
Maleimides, Acetals, Sulfhydryl Compounds, Antineoplastic Agents, Immunoconjugates, Mutation, Trastuzumab
Sponsorship
European Commission Horizon 2020 (H2020) Marie Sk?odowska-Curie actions (675007)
Identifiers
35293206, PMC8972253
External DOI: https://doi.org/10.1021/jacs.1c07675
This record's URL: https://www.repository.cam.ac.uk/handle/1810/336166
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