An Automated High Throughput Liquid Chromatography–Mass Spectrometry Process to Assess the Metabolic Stability of Drug Candidates
| العنوان: | An Automated High Throughput Liquid Chromatography–Mass Spectrometry Process to Assess the Metabolic Stability of Drug Candidates |
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| المؤلفون: | James V. Belcastro, Colleen A. McNaney, Robert Langish, Dieter M. Drexler, Adrienne A. Tymiak, Kenneth S. Santone, Serhiy Hnatyshyn, Kurt J. Edinger, Petia Shipkova, Mark Sanders, Tatyana Zvyaga, Kenneth E.J. Dickinson, Jonathan L. Josephs |
| المصدر: | ASSAY and Drug Development Technologies. 5:247-264 |
| بيانات النشر: | Mary Ann Liebert Inc, 2007. |
| سنة النشر: | 2007 |
| مصطلحات موضوعية: | Quality Control, Drug, media_common.quotation_subject, Drug Evaluation, Preclinical, Mass spectrometry, Mass Spectrometry, Liquid chromatography–mass spectrometry, Drug Discovery, Liver microsomes, NADPH-Ferrihemoprotein Reductase, media_common, Chromatography, Chemistry, Reproducibility of Results, Structural integrity, Robotics, Metabolic stability, Pharmaceutical Preparations, Data Interpretation, Statistical, Solvents, Molecular Medicine, Indicators and Reagents, Spectrophotometry, Ultraviolet, Uv detection, Software, Chromatography, Liquid |
| الوصف: | An automated high throughput process, termed the MetFast assay, is described to assess in vitro the general microsomal cytochrome P450 beta-nicotinamide adenine dinucleotide phosphate-mediated first-pass metabolic stability of potential drug candidates as a utility for pharmaceutical profiling. Utilizing robotic protocols with a multiprobe liquid handler, compounds are incubated with liver microsomes from different species. Samples are then analyzed by in-line liquid chromatography (LC)-mass spectrometry (MS) to determine the amount of compound remaining after a certain time, which allows calculation of metabolism rates. To quantitatively assess large numbers of structurally diverse compounds by LC-MS, a strategy based on an iterative two-step process was devised. Initially compounds are qualitatively analyzed by LC-ultraviolet (UV)/MS (step 1) to determine purity (UV detection) and structural integrity (MS detection). This step ensures that only correct and verified compounds with sufficient purity are being assayed to obtain reproducible high data quality. In addition, all necessary information is gathered to automatically generate specific quantitative methods for the subsequent bioanalytical analysis of metabolic stability samples by LC-UV/MS (step 2). In-house-developed, highly flexible and sophisticated data management software, termed SmartReport, is utilized for automated qualitative and quantitative LC-MS analysis set-up, data processing, and results reporting. The integration of key aspects, inherent "universal" collision-induced dissociation settings of ion trap mass spectrometers for tandem mass spectrometric scan functions utilized for compound-specific and sensitive quantitative MS methods, generic fast-LC conditions, generic MS instrument settings, and the functionality of SmartReport software resulted in an analytical process that routinely provides reproducible high-quality metabolic stability data on structurally diverse compounds. Described here is the setup of the MetFast assay, and metabolic stability data from assay validation compounds are given. |
| تدمد: | 1557-8127 1540-658X |
| DOI: | 10.1089/adt.2006.038 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d7689bed173b88b9f308efae344982c5 https://doi.org/10.1089/adt.2006.038 |
| رقم الانضمام: | edsair.doi.dedup.....d7689bed173b88b9f308efae344982c5 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15578127 1540658X |
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| DOI: | 10.1089/adt.2006.038 |