Bertini Simone, Saccomanni Giuseppe, Carlo Sara Del, Digiacomo Maria, Gargini Claudia, Piano Ilaria, Campisi Giuseppe Matteo, Ghidoni Riccardo, Macchia Marco, Manera Clementina
Dipartimento di Farmacia, Università di Pisa, Via Bonanno 6, 56126-Pisa, Italy.
Laboratorio di Biochimica e Biologia Molecolare, Dipartimento di Scienze della Salute, Via A. di Rudinì 8, 20142 Milano, Italy.
Molecules. 2017 Jul 17;22(7):1198. doi: 10.3390/molecules22071198.
In this work, we reported the application and validation of an improved high-performance liquid chromatography method coupled with a fluorimetric detector (HPLC-FL) to screen the activity of two heterocyclic derivatives reported as serine palmitoyl transferase (SPT) inhibitors. The analytical conditions were optimized in terms of the derivatization procedure, chromatographic condition, extraction procedure, and method validation according to EMEA guidelines. Once fully optimized, the method was applied to assess the SPT-inhibitory activity of the above-mentioned derivatives and of the reference inhibitor myriocin. The obtained results, expressed as a percentage of residual SPT activity, were compared to those obtained with the reference radio immune assay (RIA). The good correlation between the two types of assay demonstrated that the improved HPLC-FL method is suitable for a preliminary and rapid screening of potential SPT-inhibitors.
在本研究中，我们报道了一种改进的高效液相色谱法结合荧光检测器（HPLC-FL）用于筛选两种被报道为丝氨酸棕榈酰转移酶（SPT）抑制剂的杂环衍生物活性的应用及验证。根据欧洲药品管理局（EMEA）指南，在衍生化程序、色谱条件、提取程序和方法验证方面对分析条件进行了优化。一旦完全优化，该方法就被用于评估上述衍生物以及参考抑制剂麦角硫因的SPT抑制活性。将以残余SPT活性百分比表示的所得结果与通过参考放射免疫分析（RIA）获得的结果进行比较。两种分析方法之间的良好相关性表明，改进的HPLC-FL方法适用于潜在SPT抑制剂的初步快速筛选。
