Bioanalysis of EGFRm inhibitor osimertinib, and its glutathione cycle- and desmethyl metabolites by liquid chromatography-tandem mass spectrometry
| Autor: | Rood, J J M, van Haren, M J, Beijnen, J H, Sparidans, R W, Afd Pharmacoepi & Clinical Pharmacology, Afd Chemical Biology and Drug Discovery, Afd Pharmacology, Pharmacoepidemiology and Clinical Pharmacology, Chemical Biology and Drug Discovery, Pharmacology |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Bioanalysis
Lung Neoplasms Metabolite Proton Magnetic Resonance Spectroscopy Clinical Biochemistry Pharmaceutical Science Administration Oral 01 natural sciences Analytical Chemistry Non-small cell lung carcinoma chemistry.chemical_compound Column chromatography Liquid chromatography–mass spectrometry Tandem Mass Spectrometry Carcinoma Non-Small-Cell Lung Drug Discovery Humans Osimertinib Sulfhydryl Compounds LC-MS/MS Protein Kinase Inhibitors Spectroscopy Chromatography High Pressure Liquid Aged Aged 80 and over Acrylamides Chromatography Aniline Compounds 010405 organic chemistry 010401 analytical chemistry human plasm Glutathione Metabolism Dipeptides Desmethyl Middle Aged AZ5104 0104 chemical sciences ErbB Receptors chemistry Mutation thiol conjugates Drug Monitoring |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis, 177. Elsevier |
| ISSN: | 0731-7085 |
| Popis: | Osimertinib is a "third-generation'' oral, irreversible, tyrosine kinase inhibitor. It is used in the treatment of non-small cellular lung carcinoma and spares wild-type EGFR. Due to its reactive nature, osimertinib is, in addition to oxidative routes, metabolized through GSH coupling and subsequent further metabolism of these conjugates. The extent of the non-oxidative metabolism of osimertinib is unknown, and methods to quantify this metabolic route have not been reported yet. To gain insight into this metabolic route, a sensitive bioanalytical assay was developed for osimertinib, the active desmethyl metabolite AZ5104, and the thio-metabolites osimertinibs glutathione, cysteinylglycine, and cysteine conjugates was developed. The ease of synthesis of these metabolites was a key-part in the development of this assay. This was done through simple one-step synthesis and subsequent LC-purification. The compounds were characterized by NMR and high-resolution mass spectrometry. Sample preparation was done by a simple protein crash with acetonitrile containing the stable isotopically labeled internal standards for osimertinib and the thio-metabolites, partial evaporation of solvents, and reconstitution in eluent, followed by UHPLC-MS/MS quantification. The assay was successfully validated in a 2-2000 nM calibration range for all compounds except the glutathione metabolite, where the LLOQ was set at 6 nM due to low accuracy at 2 nM. Limited stability was observed for osimertinib, AZ5104, and the glutathione metabolite. The clinical applicability of the assay was demonstrated in samples of patients treated with 80 mg osimertinib once daily, containing all investigated compounds at detectable and quantifiable levels. |
| Databáze: | OpenAIRE |
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