Assessment of drug transporter function using fluorescent cell imaging
Kristin M Bircsak1, Christopher J Gibson, Robert W Robey
1Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey.
Current Protocols in Toxicology
|February 11, 2014
Summary
New cell-based methods efficiently detect drug-transporter interactions for breast cancer resistance protein (BCRP) and multidrug resistance protein 1 (MDR1), aiding preclinical drug development and preventing drug-drug interactions.
Area of Science:
- Pharmacology
- Biochemistry
- Drug Development
Background:
- ATP-binding cassette (ABC) proteins, such as BCRP and MDR1, actively transport diverse chemicals and mediate drug-drug interactions.
- Drug-transporter interactions can alter drug disposition, toxicity, and efficacy, impacting clinical outcomes.
- Understanding transporter roles in pharmacokinetics necessitates robust preclinical screening methods.
Purpose of the Study:
- To describe cell-based procedures for identifying functional inhibitors of BCRP and MDR1 transporters.
- To offer sensitive and rapid methods for detecting drug-transporter interactions during preclinical development.
Main Methods:
- Utilizing cell-based assays to measure fluorescent substrate accumulation in suspended cells.
- Employing an automated cell counter for sensitive and rapid intracellular fluorescence measurement.
- An alternative method using a spectrophotometer with fluorescence detection capabilities is also presented.
Main Results:
- The described methods provide convenience, sensitivity, and speed in identifying new inhibitors of BCRP and MDR1.
- Successful detection of functional inhibitors through measurement of substrate accumulation.
- Validation of automated cell counting and spectrophotometry for this application.
Conclusions:
- Cell-based assays are effective for detecting drug-transporter interactions mediated by BCRP and MDR1.
- These methods facilitate the identification of novel inhibitors, crucial for preclinical drug safety assessment.
- The described procedures enhance the ability to predict and mitigate clinical drug-drug interactions.
