the brain slice method for studying drug distribution in the cns大脑切片法研究药物分布在中枢神经系统.pdfVIP

Loryan et al. Fluids and Barriers of the CNS 2013, 10:6 FLUIDS AND BARRIERS /content/10/1/6 OF THE CNS STUDY PROTOCOL Open Access The brain slice method for studying drug distribution in the CNS 1 2 1* Irena Loryan , Markus Fridén and Margareta Hammarlund-Udenaes 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 ass