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AAPS J. 2016 May;18(3):635-46. doi: 10.1208/s12248-016-9892-3. Epub 2016 Feb 24.

Determination of Cellular Processing Rates for a Trastuzumab-Maytansinoid Antibody-Drug Conjugate (ADC) Highlights Key Parameters for ADC Design.

Author information

1
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
3
Oncology Medicinal Chemistry, Worldwide Medicinal Chemistry, Pfizer, Groton, Connecticut, USA.
4
Translational Research Group, Department of Pharmacokinetics Dynamics and Metabolism, Pfizer, Groton, Connecticut, USA.
5
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. wittrup@mit.edu.
6
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. wittrup@mit.edu.
7
Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave. 76-261D, Cambridge, Massachusetts, 02139, USA. wittrup@mit.edu.

Abstract

Antibody-drug conjugates (ADCs) are a promising class of cancer therapeutics that combine the specificity of antibodies with the cytotoxic effects of payload drugs. A quantitative understanding of how ADCs are processed intracellularly can illustrate which processing steps most influence payload delivery, thus aiding the design of more effective ADCs. In this work, we develop a kinetic model for ADC cellular processing as well as generalizable methods based on flow cytometry and fluorescence imaging to parameterize this model. A number of key processing steps are included in the model: ADC binding to its target antigen, internalization via receptor-mediated endocytosis, proteolytic degradation of the ADC, efflux of the payload out of the cell, and payload binding to its intracellular target. The model was developed with a trastuzumab-maytansinoid ADC (TM-ADC) similar to trastuzumab-emtansine (T-DM1), which is used in the clinical treatment of HER2+ breast cancer. In three high-HER2-expressing cell lines (BT-474, NCI-N87, and SK-BR-3), we report for TM-ADC half-lives for internalization of 6-14 h, degradation of 18-25 h, and efflux rate of 44-73 h. Sensitivity analysis indicates that the internalization rate and efflux rate are key parameters for determining how much payload is delivered to a cell with TM-ADC. In addition, this model describing the cellular processing of ADCs can be incorporated into larger pharmacokinetics/pharmacodynamics models, as demonstrated in the associated companion paper.

KEYWORDS:

T-DM1; antibody-drug conjugate; cellular trafficking; pharmacokinetics/pharmacodynamics; trastuzumab emtansine

PMID:
26912181
PMCID:
PMC5256610
[Available on 2017-02-24]
DOI:
10.1208/s12248-016-9892-3
[Indexed for MEDLINE]
Free PMC Article

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