Research ArticlesComparison of blood–brain barrier permeability assays: in situ brain perfusion, MDR1-MDCKII and PAMPA-BBB
Section snippets
Abbreviations:
- ACS
American Chemical Society
- AUC
area under the curve
- BBB
blood–brain barrier
- BBMEC
bovine brain microvessel endothelial cells
- BMEC
brain microvessel endothelial cells
- B/P
brain AUC (or concentration)/plasma AUC (or concentration)
- Caco-2
human colon carcinoma cell line
- Cmax
maximum concentration
- CNS
central nerve system
- MDCK
Madin-Darby canine kidney cells
- MDR
multi-drug resistance
- PAMPA
parallel artificial membrane permeability assay
- P
BBB permeability for in situ brain perfusion (×10−3 cm/s)
- Papp
apparent permeability
INTRODUCTION
Determination of brain penetration is of great importance not only for CNS drug candidates, but also for non-CNS therapeutic areas where brain penetration can cause unwanted side effects.1., 2., 3. BBB research is very challenging and active, and new findings continue to be discovered. For example, inclusion of the term “BBB” in the ACS publications has increased 100% from 2000 to 2005.3 Concerted international strategies among academia, government, and industry have been developed to address
Materials
Thirty-seven commercial drugs were obtained from Sigma (St. Louis, MO), ChemPacific (Baltimore, MD), Toronto Research Chemicals (North York, ON, Canada), Wyeth Research (Princeton, NJ). The porcine polar brain lipid (PBL) (catalog no. 141101) was from Avanti Polar Lipids, Inc. (Alabaster, AL). Universal buffer was obtained from pION Inc. (Woburn, MA). DMSO was reagent grade from Aldrich. Dodecane was from EM Science (Gibbstown, NJ). The acceptor plate was a 96-well filter plate (Multiscreen™,
RESULTS AND DISCUSSION
All the compounds listed in Ref.39 were used in the study, except for those that had weak UV absorbance, were impure or were not available at the time of the experiment. The test compounds were CNS drugs with a wide range of physico-chemical properties. This is the first large set of data published on the rate of brain penetration on marketed drugs.39 Permeability values of the 37 compounds were compared using data from three different assays: physicochemically based PAMPA-BBB, cell-based
CONCLUSIONS
The rate of brain penetration is an essential property of CNS drug candidates. In situ brain perfusion is a gold standard method for measuring BBB permeability, however, high test compound concentration in serum-free perfusion fluid can saturate Pgp efflux transport and make passive diffusion the dominant pathway. The cell-based MDR1-MDCKII assay has been widely used in the pharmaceutical industry to predict Pgp efflux transport and it is highly effective. Even though MDR1-MDCKII is an
Acknowledgements
The authors would like to thank Dr. Magid Abou-Gharbia for his support, encouragement and leadership, thank people in Pharmaceutical Profiling for their contribution and Konstantin L. Tsinman at pION Inc. for useful discussions.
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