Development of a High-Throughput Brain Slice Method for Studying Drug Distribution in the Central Nervous System
Development of a High-Throughput Brain Slice Method for Studying Drug Distribution in the Central Nervous System
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DOI：
10.1124/dmd.108.026377
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发表时间：
2009-06-01
影响因子：
3.9
通讯作者：
Hammarlund-Udenaes, Margareta
中科院分区：
文献类型：
--
作者：
Friden, Markus;Ducrozet, Frederic;Hammarlund-Udenaes, Margareta
估计中枢神经系统（CNS）中未结合药物浓度的新的、更有效的方法将通过常规方法测量的全脑组织样品中的药物量与未结合脑分布容积（V（u，脑））的体外估计相结合。虽然脑切片法是测量V（u，脑）的最可靠的体外方法，但它以前没有适应药物发现研究的需要。本研究的目的是提高该方法的通量和优化实验条件。在孵育4至5小时内，缓冲液和脑切片之间的药物平衡是基本要求。然而，在药物发现计划中，对于许多广泛结合的亲脂性化合物来说，很难满足这一要求。在本研究中，孵育容器的尺寸和搅拌模式影响平衡时间，每单位缓冲液体积的脑组织量也影响平衡时间。在线性药物浓度范围内研究V（u，脑）的盒式实验的使用增加了该方法的吞吐量。模型化合物的V（u，脑）范围为4至3000 ml。g脑（-1），和变异的来源进行了讨论。脑切片方法的优化设置允许对具有不同性质的药物（包括高度亲脂性化合物）的V（u，脑）进行精确、稳健的估计。这是在药物发现环境中实施CNS暴露相关测量的关键一步。
New, more efficient methods of estimating unbound drug concentrations in the central nervous system (CNS) combine the amount of drug in whole brain tissue samples measured by conventional methods with in vitro estimates of the unbound brain volume of distribution (V(u,brain)). Although the brain slice method is the most reliable in vitro method for measuring V(u,brain), it has not previously been adapted for the needs of drug discovery research. The aim of this study was to increase the throughput and optimize the experimental conditions of this method. Equilibrium of drug between the buffer and the brain slice within the 4 to 5 h of incubation is a fundamental requirement. However, it is difficult to meet this requirement for many of the extensively binding, lipophilic compounds in drug discovery programs. In this study, the dimensions of the incubation vessel and mode of stirring influenced the equilibration time, as did the amount of brain tissue per unit of buffer volume. The use of casette experiments for investigating V(u,brain) in a linear drug concentration range increased the throughput of the method. The V(u,brain) for the model compounds ranged from 4 to 3000 ml . g brain(-1), and the sources of variability are discussed. The optimized setup of the brain slice method allows precise, robust estimation of V(u,brain) for drugs with diverse properties, including highly lipophilic compounds. This is a critical step forward for the implementation of relevant measurements of CNS exposure in the drug discovery setting.