Harmonised high throughput microsomal stability assay
Harmonised high throughput microsomal stability assay
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DOI：
10.1016/j.vascn.2016.10.006
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发表时间：
2017-03-01
影响因子：
1.9
通讯作者：
Riley, Robert J.
中科院分区：
文献类型：
--
作者：
Williamson, Beth;Wilson, Claire;Riley, Robert J.
引言：从体外试验和临床前数据预测人体药代动力学是药物发现的一个组成部分。体外稳定性代谢研究可通过纳入生物比例因子来提供体内肝固有清除率的估计值。许多实验室都有个性化的稳定性方案，包括标记化合物，并采用了QC标准和测定限度，以确保数据的完整性。合同研究组织（CRO）为学术和制药生物技术机构提供综合药物发现支持，以推进其内部项目。大多数这些客户已经建立了相关的标准，以确保内部和外部labels.Methods之间的数据一致性的内部协议：在这项研究中，众多的分析变量被浓缩成一个协调的分析格式和一系列的化合物与不同的物理化学性质进行了评估。该协议是不同的以下属性：缓冲液，微粒体浓度和物种strain.Results：比较人的批次在体外克林特之间的传统和巩固的检测格式显示出良好的相关性，没有显着差异。菌株之间存在明显的相关性。各种属菌株之间的体外固有清除率解释一致。使用严格的体外肝固有清除率值（150 μ L/min/mg蛋白质），在不同条件下，如测定变量，人批次，小鼠和大鼠品系的比较显示>80% agreement.Discussion：开发了一种高通量测定法，可以使用小鼠，大鼠和人肝微粒体（合并测定）同时测量克林特。通过将所有可能的变量压缩到一种测定格式中，在头对头测试期间获得了一致的数据。(C)2016 Elsevier Inc. All rights reserved.
Introduction: Prediction of human pharmacokinetics from in vitro assays and pre-clinical data is an integral part of drug discovery. in vitro stability metabolic studies can provide an estimate of in vivo hepatic intrinsic clearance through inclusion of biological scaling factors. Many labs have personalised stability protocols including marker compounds and have adopted QC criteria and assay limits to ensure data integrity. Contract research organisanons (CRO's) provide integrated drug discovery support to academic and pharmaceutical biotechnology institunons to progress their in-house projects. The majority of these clients have established in-house protocols with associated criteria to ensure data consistency between in-house and external labs.Methods: In this study, numerous assay variables were condensed into one harmonised assay format and a range of compounds with diverse physicochemical properties were evaluated. The protocols were diverse with respect to the following attributes: buffer, microsomal concentration and species strain.Results: Comparison of human lots in vitro CLint between the traditional and consolidated assay formats showed a good correlation with no significant difference. A clear relationship was demonstrated between strains. Interpretation of in vitro intrinsic clearance between the strains for each species was consistent. Using strict classes of in vitro hepatic intrinsic clearance values ( 150 mu L/min/mg protein) comparisons across different conditions such as, assay variables, human lots, mouse and rat strains showed >80% agreement.Discussion: A high throughput assay was developed that enables the simultaneous measurement of CLint using mouse, rat and human hepatic microsomes (consolidated assay). By condensing all possible variables into one assay format, consistent data were obtained during head to head tests. (C) 2016 Elsevier Inc. All rights reserved.