Schouten Wietse M, van Bocxlaer Katrien, Rosing Hilde, Huitema Alwin D R, Beijnen Jos H, Kratz Jadel M, Mowbray Charles E, Dorlo Thomas P C
Department of Pharmacy and Pharmacology, Antoni van Leeuwenhoek/The Netherlands Cancer Institute, Amsterdam, the Netherlands.
Skin Research Centre, Hull York Medical School, York Biomedical Research Institute, University of York, York, UK.
Biomed Chromatogr. 2025 Aug;39(8):e70158. doi: 10.1002/bmc.70158.
Understanding the target site pharmacokinetics (PK) of the nitroimidazole analog DNDI-0690, a potential drug for the neglected parasitic disease leishmaniasis, is important due to the diversity of infected tissue sites and potential drug penetration variability. An ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for quantifying DNDI-0690 in murine biomatrices (plasma, liver, spleen, skin, and skin microdialysate). The method used three protein precipitation sample preparation procedures, tailored for different biomatrices, utilizing a surrogate biomatrix approach. Murine tissues were enzymatically homogenized with a Collagenase A mixture. Chromatographic detection was performed on a C18 column using gradient elution, coupled to a QTRAP6500 quadrupole MS, operating in positive ionization mode. The method demonstrated accurate and precise quantification of all murine biomatrices on the surrogate biomatrix calibration standards, with a high and reproducible total recovery ranging from 75.9% to 94.2% (CV% ≤ 2.5%). Matrix interferences were mitigated with a deuterated internal standard. Stability experiments demonstrated that DNDI-0690 remained stable in all biomatrices under various conditions. This validated UHPLC-MS/MS method was successfully used to quantify DNDI-0690 in a target site murine infection model, demonstrating its suitability for future target site PK studies involving DNDI-0690.
了解硝基咪唑类似物DNDI - 0690的靶位点药代动力学（PK）很重要，它是一种用于治疗被忽视的寄生虫病利什曼病的潜在药物，因为感染组织部位具有多样性且药物渗透可能存在变异性。开发并验证了一种超高效液相色谱 - 串联质谱（UHPLC - MS/MS）方法，用于定量小鼠生物基质（血浆、肝脏、脾脏、皮肤和皮肤微透析液）中的DNDI - 0690。该方法采用了三种针对不同生物基质定制的蛋白质沉淀样品制备程序，利用替代生物基质方法。用胶原酶A混合物对小鼠组织进行酶解匀浆。在C18柱上进行梯度洗脱色谱检测，与在正离子模式下运行的QTRAP6500四极杆质谱联用。该方法在替代生物基质校准标准品上对所有小鼠生物基质进行了准确且精确的定量，总回收率高且可重复，范围为75.9%至94.2%（CV%≤2.5%）。用氘代内标减轻了基质干扰。稳定性实验表明，DNDI - 0690在各种条件下在所有生物基质中均保持稳定。这种经过验证的UHPLC - MS/MS方法成功用于在靶位点小鼠感染模型中定量DNDI - 0690，证明了其适用于未来涉及DNDI - 0690的靶位点PK研究。
