title: A rapid, accurate and robust UHPLC-MS/MS method for quantitative determination of BMS-927711, a CGRP receptor antagonist, in plasma in support of non-clinical toxicokinetic studies.
abstract: BMS-927711 is a calcitonin gene-related peptide (CGRP) receptor antagonist that is being developed for the treatment of migraine. A rapid, accurate and robust assay was developed and validated for the quantitation of BMS-927711 in rat, monkey, rabbit and mouse plasma using ultra high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). A simplified method screening strategy was utilized that included a liquid-liquid extraction (LLE) methodology and eleven LC columns (ten sub-2 μm UHPLC columns and one 2.6 μm HPLC column) for screening with emphasis on the removal of phospholipids, avoidance of metabolite interference and ruggedness of LC conditions. A stable isotope labeled [(13)C2, D4]-BMS-927711 was used as the internal standard, and 50 μL of plasma samples were used for extraction by automated LLE with methyl tert-butyl ether (MTBE) in 96-well format. Chromatographic separation was achieved with an isocratic elution and a gradient column wash on a Waters Acuity UPLC(®) BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with run time of 3.7 min. Positive electrospray ionization was performed using selected reaction monitoring (SRM) with transitions of m/z 535>256 for BMS-927711 and m/z 541>256 for [(13)C2, D4]-BMS-927711. The standard curve, which ranged from 3.00 to 3000 ng/mL for BMS-927711, was fitted to a 1/x(2) weighted linear regression model. The intra-assay precision was within 5.2% CV, inter-assay precision was within 5.9% CV, and the assay accuracy was within ±5.2% deviation (%Dev) of the nominal values in all the species. The stability of an N-carbamoyl glucuronide metabolite was carefully investigated, and the conversion of this metabolite to BMS-927711 was minimal and manageable without a stabilization procedure. The method was successfully applied to multiple non-clinical toxicokinetic studies in different species in support of the investigative new drug (IND) filing.
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb Company, Princeton, NJ 08543, USA. naiyu.zheng@bms.com
authors.forename: Naiyu
authors.initials: N
authors.lastname: Zheng
authors.forename: Jianing
authors.initials: J
authors.lastname: Zeng
authors.forename: Billy
authors.initials: B
authors.lastname: Akinsanya
authors.forename: Adela
authors.initials: A
authors.lastname: Buzescu
authors.forename: Yuan-Qing
authors.initials: YQ
authors.lastname: Xia
authors.forename: Van
authors.initials: V
authors.lastname: Ly
authors.forename: Kevin
authors.initials: K
authors.lastname: Trouba
authors.forename: Qianping
authors.initials: Q
authors.lastname: Peng
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Aubry
authors.forename: Mark E
authors.initials: ME
authors.lastname: Arnold
mesh_terms: D000818:Animals; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D020719:Glucuronides / Q000737:chemistry; D000882:Haplorhini; D059625:Liquid-Liquid Extraction / Q000379:methods; D051379:Mice; D010880:Piperidines / Q000097:blood* / Q000737:chemistry*; D011725:Pyridines / Q000097:blood* / Q000737:chemistry*; D011817:Rabbits; D051381:Rats; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D020719:Glucuronides; D010880:Piperidines; D011725:Pyridines; C578443:rimegepant sulfate