Masimirembwa C M, Thompson R, Andersson T B
Department of DMPK & Bioanalytical Chemistry, AstraZeneca, R&D Mölndal, S431 83 Mölndal, Sweden.
Comb Chem High Throughput Screen. 2001 May;4(3):245-63. doi: 10.2174/1386207013331101.
Drug metabolism can have profound effects on the pharmacological and toxicological profile of therapeutic agents. In the pharmaceutical industry, many in vitro techniques are in place or under development to screen and optimize compounds for favorable metabolic properties in the drug discovery phase. These in vitro technologies are meant to address important issues such as: (1) is the compound a potent inhibitor of drug metabolising enzymes (DMEs)? (2) does the compound induce the expression of DMEs? (3) how labile is the compound to metabolic degradation? (4) which specific enzyme(s) is responsible for the compound's biotransformation? and (5) to which metabolites is the compound metabolized? Answers to these questions provide a basis for judging whether a compound is likely to have acceptable pharmacokinetic properties in vivo. To address these issues on the increasing number of compounds inundating the drug discovery programs, high throughput assays are essential. A combination of biochemical advances in the understanding of the function and regulation of DMEs (in particular, cytochromes P450, CYPs) and automated analytical technologies are revolutionizing drug metabolism research. Automated LC-MS based metabolic stability, fluorescence, radiometric and LC-MS based CYP inhibition assays are now in routine use. Automatible models for studying CYP induction based on enzyme activity, quantitative RT-PCR and reporter gene systems are being developed. We will review the utility and limitations of these HTS approaches and highlight on-going developments and emerging technologies to answer metabolism questions at the different stages of the drug discovery process.
药物代谢可对治疗药物的药理和毒理学特性产生深远影响。在制药行业，许多体外技术已得到应用或正在开发中，以便在药物发现阶段筛选和优化具有良好代谢特性的化合物。这些体外技术旨在解决重要问题，例如：（1）该化合物是否为药物代谢酶（DME）的强效抑制剂？（2）该化合物是否诱导DME的表达？（3）该化合物对代谢降解的稳定性如何？（4）哪种特定酶负责该化合物的生物转化？以及（5）该化合物代谢成哪些代谢物？对这些问题的回答为判断化合物在体内是否可能具有可接受的药代动力学特性提供了依据。为了解决涌入药物发现项目的化合物数量不断增加所带来的这些问题，高通量分析至关重要。对DME（特别是细胞色素P450，CYPs）功能和调节的理解方面的生化进展与自动化分析技术相结合，正在彻底改变药物代谢研究。基于自动化液相色谱 - 质谱的代谢稳定性、荧光、放射性测定以及基于液相色谱 - 质谱的CYP抑制测定现在已常规使用。基于酶活性、定量逆转录 - 聚合酶链反应和报告基因系统的用于研究CYP诱导的可自动化模型正在开发中。我们将回顾这些高通量方法的实用性和局限性，并重点介绍正在进行的发展和新兴技术，以回答药物发现过程不同阶段的代谢问题。
