Drug Disposition Eli Lilly and Company, Indianapolis, USA.
J Pharm Pharm Sci. 2016 Oct-Dec;19(4):496-510. doi: 10.18433/J3NK60.
Current practices applied to mouse pharmacokinetic (PK) studies often use large numbers of animals with sporadic or composite sampling that inadequately describe PK profiles. The purpose of this work was to evaluate and optimize blood microsampling techniques coupled with dried blood spot (DBS) and LC-MS/MS analysis to generate reliable PK data in mice. In addition, the feasibility of cross-over designs was assessed and recommendations are presented.
The work describes a comprehensive evaluation of five blood microsampling techniques (tail clip, tail vein with needle hub, submandibular, retro-orbital, and saphenous bleeding) in CD-1 mice. The feasibility of blood sampling was evaluated based on animal observations, ease of bleeding, and ability to collect serial samples. Methotrexate, gemfibrozil and glipizide were used as test compounds and were dosed either orally or intravenously, followed by DBS collection and LC-MS/MS analysis to compare PK with various bleeding methods.
Submandibular and retro-orbital methods that required non-serial blood collections did not allow for inter-animal variability assessments and resulted in poorly described absorption and distribution kinetics. The submandibular and tail vein with needle-hub methods were the least favorable from a technical feasibility perspective. Serial bleeding was possible with cannulated animals or saphenous bleeding in non-cannulated animals.
Of the methods that allowed serial sampling, the saphenous method when executed as described in this report, was most practical, reproducible and provided for assessment of inter-animal variability. It enabled the collection of complete exposure profiles from a single mouse and the conduct of an intravenous/oral cross-over study design. This methodology can be used routinely, it promotes the 3Rs principles by achieving reductions in the number of animals used, decreased restraints and animal stress, and improved the quality of data obtained in mouse PK studies. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.
目前应用于小鼠药代动力学（PK）研究的实践通常使用大量具有零星或复合采样的动物，这些采样方法不能充分描述 PK 曲线。本研究旨在评估和优化血液微采样技术，并结合干血斑（DBS）和 LC-MS/MS 分析，以生成小鼠可靠的 PK 数据。此外，还评估了交叉设计的可行性，并提出了建议。
本研究全面评估了 CD-1 小鼠五种血液微采样技术（尾夹、带针座的尾静脉、颌下、眶后和隐静脉采血）。根据动物观察、采血难易程度和连续采样能力，评估采血的可行性。甲氨蝶呤、吉非贝齐和格列吡嗪被用作测试化合物，分别经口或静脉给药，然后采集 DBS 并进行 LC-MS/MS 分析，以比较不同采血方法的 PK 数据。
需要非连续采血的颌下和眶后方法不能评估动物间的变异性，导致吸收和分布动力学描述不佳。从技术可行性的角度来看，颌下和带针座的尾静脉方法最不理想。使用套管动物进行连续采血或在非套管动物中进行隐静脉采血是可行的。
在允许连续采样的方法中，如本报告所述执行的隐静脉采血方法最为实用、可重现，并且可以评估动物间的变异性。它能够从单个小鼠中采集完整的暴露曲线，并进行静脉/口服交叉设计研究。这种方法可以常规使用，通过减少使用的动物数量、减少束缚和动物应激，以及提高在小鼠 PK 研究中获得的数据质量，促进了 3R 原则。本文接受 POST-PUBLICATION REVIEW。注册读者（请参阅“读者须知”）可以在本期内容的页面上点击 ABSTRACT 进行评论。
