A Reactive Antibody Platform for One-Step Production of Antibody–Drug Conjugates through a Diels–Alder Reaction with Maleimide
| Autor: | Fengying Huang, Balakumar Vijayakrishnan, Chacko Chakiath, Shenlan Mao, Daniel Lemen, Christine Fazenbaker, Marcello Marelli, Javier Read de Alaniz, Neki V. Patel, Lauren Adams, Philip Howard, Haihong Zhong, Wenshu Xu, R. James Christie, André H. St. Amant, Jay Harper, Changshou Gao, Herren Wu, Jia Lin |
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| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Immunoconjugates Cell Survival Protein Conformation Biomedical Engineering Pharmaceutical Science Bioengineering CHO Cells 02 engineering and technology 01 natural sciences Adduct Maleimides chemistry.chemical_compound Cricetulus Animals Humans Spiro Compounds Maleimide Diels–Alder reaction Pharmacology chemistry.chemical_classification Bioconjugation Cycloaddition Reaction 010405 organic chemistry Organic Chemistry 021001 nanoscience & nanotechnology Combinatorial chemistry Cycloaddition 3. Good health 0104 chemical sciences Amino acid chemistry Covalent bond PC-3 Cells 0210 nano-technology Biotechnology Conjugate |
| Zdroj: | Bioconjugate Chemistry. 30:2340-2348 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/acs.bioconjchem.9b00436 |
| Popis: | The normal electron-demand Diels-Alder (DA) cycloaddition is a classic transformation routinely used in synthesis; however, applications in biological systems are limited. Here, we report a spiro[2.4]hepta-4,6-diene-containing noncanonical amino acid (SCpHK) capable of efficient incorporation into antibodies and subsequent coupling with maleimide via a DA reaction. SCpHK was stable throughout protein expression in mammalian cells and enabled covalent attachment of maleimide drug-linkers yielding DA antibody-drug conjugates (DA-ADCs) with nearly quantitative conversion in a one-step process. The uncatalyzed DA reaction between SCpHK and maleimide in aqueous buffer was rapid (1.8-5.4 M-1 s-1), and the antibody-drug adduct was stable in rat serum for at least 1 week at 37 °C. Anti-EphA2 DA-ADCs containing AZ1508 or SG3249 maleimide drug-linkers were potent inhibitors of tumor growth in PC3 tumor models in vivo. The DA bioconjugation strategy described here represents a simple method to produce site-specific and stable ADCs with maleimide drug-linkers. |
| Databáze: | OpenAIRE |
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