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Open Access Highly Accessed Study protocol

The brain slice method for studying drug distribution in the CNS

Irena Loryan1, Markus Fridén2 and Margareta Hammarlund-Udenaes1*

Author Affiliations

1 Department of Pharmaceutical Biosciences, Translational PKPD Research Group, Uppsala University, Associate member of SciLife Lab, Box 591, Uppsala, SE-75124, Sweden

2 AstraZeneca Research and Development, Respiratory and Inflammatory Innovative Medicines, Department of Drug Metabolism and Pharmacokinetics, Mölndal, Sweden

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Fluids and Barriers of the CNS 2013, 10:6  doi:10.1186/2045-8118-10-6

Published: 21 January 2013

Abstract

The high-throughput brain slice method is a precise and robust technique for estimating the overall uptake of drugs into brain tissue through determination of the unbound volume of distribution in the brain (Vu,brain; ml·g brain-1). Vu,brain describes the relationship between the total drug concentration in the brain and the concentration of unbound drug in the brain interstitial fluid, regardless of blood–brain barrier function. The brain slice method is more physiologically based than the brain homogenate method with respect to the assessment of drug distribution in the brain because the cell-cell interactions, pH gradients and active transport systems are all conserved. The method provides information that is directly relevant to issues such as nonspecific binding to brain tissue, lysosomal trapping, and active uptake into the cells. For these reasons, the brain slice method is recommended for estimation of target-site pharmacokinetics in the early drug discovery process and fundamental pharmacological studies. This article provides a detailed protocol for the rat and mouse brain slice methods, with the aim of enabling simple, cost-effective profiling of compounds with diverse physicochemical properties. The procedure for assessing the viability of the brain slices after the 5 h incubation period is also described. The results are interpreted for a set of compounds covering a wide range of physicochemical properties and various pharmacological targets. Application of the method for evaluating the unbound intracellular-to-extracellular concentration ratio (Kp,uu,cell) and the unbound brain-to-plasma concentration ratio (Kp,uu,brain) is discussed.

Keywords:
Brain slice method; Unbound volume of distribution in the brain; Neuropharmacokinetics; Drug discovery; High-throughput screening