Quantification of Proteins Involved in Drug Metabolism and Disposition in the Human Liver Using Label-Free Global Proteomics
Quantification of Proteins Involved in Drug Metabolism and Disposition in the Human Liver Using Label-Free Global Proteomics
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DOI：
10.1021/acs.molpharmaceut.8b00941
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发表时间：
2019-02-01
影响因子：
4.9
通讯作者：
Barber, Jill
中科院分区：
文献类型：
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作者：
Couto, Narciso;Al-Majdoub, Zubida M.;Barber, Jill
目前迫切需要（在FDA指南2018中得到认可）优化给予患者的药物剂量，以实现最大药物疗效和有限毒性（精确给药），这可以通过定量系统药理学（QSP）模型来促进。准确定量参与药物清除的蛋白质对于建立和改进任何目标人群的QSP模型至关重要。本文应用无标记蛋白质组学技术对23例人肝微粒体中188种酶和66种转运蛋白进行了定量分析，其中包括17种细胞色素P450（CYP）、10种UDP-葡萄糖醛酸转移酶样品（UGT）、7种ATP结合盒（ABC）转运蛋白和11种溶质载体样品（SLC）转运蛋白;这些蛋白质中的六种首次被定量。该方法允许定量数千种蛋白质，允许估计样品纯度和理解蛋白质表达的全球模式。在可获得的情况下，与靶向定量和酶活性数据总体一致。评估了性别、年龄、基因型和BMI对酶和转运蛋白表达的影响。随着BMI的增加，观察到酶和转运蛋白的表达减少，但老年供体具有较高BMI的趋势可能混淆了这一结果。然而，基因型对酶表达的影响是明确的，与CYP 3AS *1/*3基因型相比，其主要无活性的 *3/*3对应物表达高16倍。尽管无标记方法需要复杂、耗时的数据分析，但无标记方法的优势使其成为一种有价值的方法，可以在药理学和毒理学模型中同时填充目标患者的广泛系统参数。
There is an urgent need (recognized in FDA guidance, 2018) to optimize the dose of medicines given to patients for maximal drug efficacy and limited toxicity (precision dosing), which can be facilitated by quantitative systems pharmacology (QSP) models. Accurate quantification of proteins involved in drug clearance is essential to build and improve QSP models for any target population. Here we describe application of label-free proteomics in microsomes from 23 human livers to simultaneously quantify 188 enzymes and 66 transporters involved in xenobiotic disposition, including 17 cytochrome P450s (CYPs), 10 UDP-glucuronosyltransferases Samples (UGTs), 7 ATP-binding cassette (ABC) transporters, and 11 solute carrier Sample (SLC) transporters; six of these proteins are quantified for the first time. The methodology allowed quantification of thousands of proteins, allowing estimation of sample purity and understanding of global patterns of protein expression. There was overall good agreement with targeted quantification and enzyme activity data, where this was available. The effects of sex, age, genotype, and BMI on enzyme and transporter expression were assessed. Decreased expression of enzymes and transporters with increasing BMI was observed, but a tendency for older donors to have higher BMIs may have confounded this result. The effect of genotype on enzymes expression was, however, clear-cut, with CYP3AS*1/*3 genotype expressed 16-fold higher compared with its mostly inactive *3/*3 counterpart. Despite the complex, time-consuming data analysis required for label-free methodology, the advantages of the label-free method make it a valuable approach to populate a broad range of system parameters simultaneously for target patients within pharmacology and toxicology models.