Hart Robert, Karnik Shane, Van Sickle Kathy, Mullin Mark, Naderer Odin, Tippin Tim, Dunn John
Clinical Pharmacology and Translational Medicine, Chimerix, Inc., Durham, NC 27705, USA.
Laboratory operations, Colorado, Aliri Bioanalysis, Colorado Springs, CO 80907, USA.
Bioanalysis. 2024;16(21-22):1179-1187. doi: 10.1080/17576180.2024.2418245. Epub 2024 Oct 30.
Brincidofovir has been approved by the US FDA for the treatment of smallpox via the "Animal Rule". The active moiety, cidofovir diphosphate (CDV-PP), was measurable in human peripheral blood mononuclear cells (PBMCs), but quantitation in animals was challenging given their limited blood volume. The aim of this study was to optimize PBMC isolation from rabbit and mouse blood to allow quantitation of CDV-PP. PBMC isolation methods using various separation media were evaluated using blood from naive and brincidofovir-dosed animals. CDV-PP was quantified using liquid chromatography tandem mass spectrometry. PBMC isolation yields were increased by >fourfold compared with yields obtained using human methods, allowing cross-species exposure comparisons.
布立昔洛韦已通过“动物规则”获得美国 FDA 批准，用于天花的治疗。活性部分，更昔洛韦二磷酸（CDV-PP），可在人体外周血单核细胞（PBMCs）中检测到，但由于动物的血液体积有限，定量检测具有挑战性。本研究旨在优化从兔和鼠血中分离 PBMC 的方法，以允许定量检测 CDV-PP。使用来自未处理和布立昔洛韦处理动物的血液评估了使用各种分离介质的 PBMC 分离方法。使用液相色谱串联质谱法定量 CDV-PP。与使用人体方法获得的产量相比，PBMC 分离产量增加了四倍以上，从而允许进行跨物种暴露比较。
