BD-Application of BCRP, BSEP and MRP2 Transporter Vesicles in Drug Development Studies

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ABC Transporter Membrane Vesicles Available from BD
BD GentestTM ABC Transporter Membrane Vesicles
ABC: ATP-Binding Cassette Family of Transporters
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Uptake Assay in ABC Transporter Vesicles
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Assessment of additional transporter-dependent drug disposition and drug interactions is now routinely expected by regulatory agencies Recent consensus by the USFDA and industry representatives has outlined guidance for assessment of ABC and SLC transporter-mediated interactions during drug development
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Role of Transporters in PK/PD
• Efflux of drug from various tissues
– BCRP (rosuvastatin) – BSEP (vinblastine)
• Vectorial transport of drugs into and out of hepatocytes
Disadvantages
• • Potential that extremely high permeability compounds could generate high background (in control condition) Typically requires radiolabeled test articles for substrate ID (non-labeled compounds can be assayed as potential inhibitors of radiolabeled probe substrates)
Pravastatin Portal vein Uptake by unconfirmed transporter Liver Uptake by OATP1B1 Bile Efflux by MRP2
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Importance of In Vitro Transporter Assays in Drug Development
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Giacomini, Huang, Tweedie et al, Nature Reviews Drug Discovery (2010) 9:215-236
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Guidance and Position Papers
• Tucker et al. Optimizing drug development: strategies to assess drug metabolism/transporter interaction potential – toward a consensus. Basel Conference 2001, Sponsored by FDA, EUFEPS, AAPS:
Application of BCRP, BSEP and MRP2 Transporter Vesicles in Drug Development Studies
Lisa Fox
June 2, 2010
Topics
• Drug-transporter interaction in drug development • ABC Transporter membrane vesicles • Uptake and inhibition kinetics in vesicles • Drug development study designs • Other transporter applications
– – – – Clin Pharmacol Ther (2001), 70:103-14, Br J Clin Pharmacol (2001), 52:107-17, Eur J Pharm Sci (2001), 13:417-28, Pharm Res (2001), 18:1071-80
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Regulatory Expectations and Guidance
• FDA Guidance (2006)* outlines recommendations for assessing drug interactions with human MDR1 (ABCB1)
* FDA Draft Guidance for Industry (Sept 2006) Drug Interaction Studies – Study Design, Data Analysis and Implications for Dosing and Labeling (/cder/guidance/index.htm)
ATP-dependent uptake of drugs into vesicles via “inside-out” efflux transporters
Inside Out
ATP
Drug
ADP + P
Vesicles prepared from baculovirus/Sf9 insect cell expression system Efflux transporter is in an “inside-out” or reverse orientation on the sphere Orientation allows substrates to be taken up by the transporter into the vesicles 96-well reaction plate filter plate via cell harvester/vacuum manifold LSC Measures uptake activity of test articles in the absence and presence of known inhibitors
Zhang et al. (2006) Scientific Perspectives on Drug Transporters and Their Role in Drug Interactions. Mol Pharm 3:62-69 FDA DRAFT Guidance for Industry (Sept 2006) Drug Interaction Studies – Study Design, Data Analysis and Implications for Dosing and Labeling (/cder/guidance/index.htm) Huang et al. (2008) New Era in Drug Interaction Evaluation: US FDA Update on CYP Enzymes, Transporters, and the Guidance Process. J Clin Pharmacol 48:662-670 Zhang et al. (2008) A Regulatory Viewpoint on Transporter-based Drug Interactions. Xenobiotica 38:709-724 Giacomini, et. al. Membrane Transporters in Drug Development – The International Transporter Consortium Nature Reviews: Drug Discovery (Mar 2010) 9:215-236 EMEA DRAFT: European Medicines Agency Guideline on the Investigation of Drug Interactions, (Apr 2010) CPMP/EWP/560/95/Rev. 1 – Corr.
– SLC Transporters
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Importance of BCRP, BSEP and MRP2 in Drug Development
Consensus of the International Transporter Consortium: Giacomini, Huang, Tweedie et al, Nature Reviews Drug Discovery (2010) 9:215-236
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Time- and Temperature-Dependence
Characterization data with human MRP2 membrane vesicles demonstrating linear time-dependence and effect of temperature on transport activity
• Understand function of transporters in the absorption, distribution and elimination of drug candidates • Evaluate and improve drug bioavailability (transporter-SAR) • Predict and avoid transporter involved DDIs • Explain/predict pharmacokinetic data (e.g. non-proportional PK, excretion of unchanged drug) • To provide information in support of IND or NDA filings addressing potential transporter-mediated drugVesicle Model