title: Validation of high-performance liquid chromatography-tandem mass spectrometry assays quantifying omacetaxine mepesuccinate and its 4'‑des-methyl and cephalotaxine metabolites in human plasma and urine.
abstract: Omacetaxine mepesuccinate (hereafter called omacetaxine) is a modified cephalotaxine and is registered (Synribo(®)) for the treatment of adult patients with chronic myeloid leukemia (CML) with resistance and/or intolerance to two or more tyrosine kinase inhibitors (TKIs). To evaluate the pharmacokinetics of omacetaxine, sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays for the quantification of omacetaxine and its inactive 4'-des-methyl (4'-DMHHT) and cephalotaxine metabolites in human plasma and urine were developed and validated. Since omacetaxine is mainly metabolised by esterases, the plasma samples were immediately stabilised after collection with an esterase inhibitor and stored at a nominal temperature of -80°C. Urine samples were stored at -80°C immediately after collection. Protein precipitation was applied as the sample pretreatment method for the plasma samples, and urine samples were processed using solid-phase extraction (SPE). For both assays, the dried and reconstituted extracts were injected on a XBridge BEH Phenyl column for analysis of all analytes. Gradient elution was applied with 0.1% formic acid in water and methanol as mobile phases. Analytes were ionised using a turbospray ionisation source in positive mode and detected with a triple quadrupole mass spectrometer. The validated plasma assay quantifies all analytes in the concentration range of 0.1-100ng/mL and the urine assay in the range of 0.1-50ng/mL. At all concentrations, the accuracies were within ±15% of the nominal concentrations and precisions were ≤15%. The developed methods have successfully been applied in a human mass balance study of omacetaxine.
authors: Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands. Electronic address: Cynthia.Nijenhuis@slz.nl.
authors: C M
authors: CM
authors: Nijenhuis
authors: Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors: L
authors: L
authors: Lucas
authors: Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors: H
authors: H
authors: Rosing
authors: Division of Clinical Pharmacology, Department of Medical Oncology, The Netherlands Cancer Institute, Amsterdam, The Netherlands; Division of Pharmacoepidemiology and Clinical Pharmacology, Faculty of Science, Department of Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors: J H M
authors: JH
authors: Schellens
authors: Department of Pharmacy & Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, The Netherlands; Division of Pharmacoepidemiology and Clinical Pharmacology, Faculty of Science, Department of Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors: J H
authors: JH
authors: Beijnen
authors: Nonclinical DMPK, Teva Branded Pharmaceutical R&D, Inc., West Chester, PA, USA.
authors: S H
authors: SH
authors: Gorman
authors: Quintiles BioSciences, Inc., Ithaca, NY, USA.
authors: S M
authors: SM
authors: Burke
authors: Quintiles BioSciences, Inc., Ithaca, NY, USA.
authors: D A
authors: DA
authors: Campbell
authors: Quintiles BioSciences, Inc., Ithaca, NY, USA.
authors: M W
authors: MW
authors: Chapple
authors: Quintiles BioSciences, Inc., Ithaca, NY, USA.
authors: T H
authors: TH
authors: Yousey
authors: Quintiles BioSciences, Inc., Ithaca, NY, USA.
authors: D E
authors: DE
authors: Mulvana
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000097:blood* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006248:Harringtonines / Q000097:blood* / Q000652:urine*; D000077863:Homoharringtonine; D006801:Humans; D057230:Limit of Detection; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D006248:Harringtonines; D000077863:Homoharringtonine
keywords: 4′-Des-methylhomoharringtonine; Cephalotaxine; GLP; HPLC–MS/MS; Omacetaxine mepesuccinate