Cathepsin B Cleavage of vcMMAE-Based Antibody–Drug Conjugate Is Not Drug Location or Monoclonal Antibody Carrier Specific
| Autor: | Li Yi, Benson Gikanga, Hung-Wei Chih, Nia Adeniji, Thomas W. Patapoff |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Drug Antibody-drug conjugate Immunoconjugates media_common.quotation_subject Biomedical Engineering Pharmaceutical Science Bioengineering Conjugated system Bioinformatics Peptide Mapping Cathepsin B Accessible surface area 03 medical and health sciences 0302 clinical medicine media_common Pharmacology Cathepsin Chemistry Organic Chemistry Antibodies Monoclonal Small molecule body regions Kinetics 030104 developmental biology 030220 oncology & carcinogenesis Proteolysis Biophysics Oligopeptides Biotechnology Conjugate |
| Zdroj: | Bioconjugate Chemistry. 27:1040-1049 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/acs.bioconjchem.6b00055 |
| Popis: | Antibody-drug conjugates (ADCs) require thorough characterization and understanding of product quality attributes. The framework of many ADCs comprises one molecule of antibody that is usually conjugated with multiple drug molecules at various locations. It is unknown whether the drug release rate from the ADC is dependent on drug location, and/or local environment, dictated by the sequence and structure of the antibody carrier. This study addresses these issues with valine-citrulline-monomethylauristatin E (vc-MMAE)-based ADC molecules conjugated at reduced disulfide bonds, by evaluating the cathepsin B catalyzed drug release rate of ADC molecules with different drug distributions or antibody carriers. MMAE drug release rates at different locations on ADC I were compared to evaluate the impact of drug location. No difference in rates was observed for drug released from the V(H), V(L), or C(H)2 domains of ADC I. Furthermore, four vc-MMAE ADC molecules were chosen as substrates for cathepsin B for evaluation of Michaelis-Menten parameters. There was no significant difference in K(M) or k(cat) values, suggesting that different sequences of the antibody carrier do not result in different drug release rates. Comparison between ADCs and small molecules containing vc-MMAE moieties as substrates for cathepsin B suggests that the presence of IgG1 antibody carrier, regardless of its bulkiness, does not impact drug release rate. Finally, a molecular dynamics simulation on ADC II revealed that the val-cit moiety at each of the eight possible conjugation sites was, on average, solvent accessible over 50% of its maximum solvent accessible surface area (SASA) during a 500 ns trajectory. Combined, these results suggest that the cathepsin cleavage sites for conjugated drugs are exposed enough for the enzyme to access and that the drug release rate is rather independent of drug location or monoclonal antibody carrier. Therefore, the distribution of drug conjugation at different sites is not a critical parameter to control in manufacturing of the vc-MMAE-based ADC conjugated at reduced disulfide bonds. |
| Databáze: | OpenAIRE |
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