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.
Application of An Improved HPLC-FL Method to Screen Serine Palmitoyl Transferase Inhibitors / S. Bertini, G. Saccomanni, S.D. Carlo, M. Digiacomo, C. Gargini, I. Piano, G.M. Campisi, R. Ghidoni, M. Macchia, C. Manera. - In: MOLECULES. - ISSN 1420-3049. - 22:7(2017 Jul).
|Titolo:||Application of An Improved HPLC-FL Method to Screen Serine Palmitoyl Transferase Inhibitors|
|Parole Chiave:||enzymatic assay; HPLC; screening; serine palmitoyl transferase; SPT; dose-response relationship, drug; enzyme inhibitors; HEK293 cells; humans; magnetic resonance spectroscopy; molecular structure; reproducibility of results; serine C-palmitoyltransferase; substrate specificity; chromatography, high pressure liquid; fluorometry; organic chemistry|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
|Data di pubblicazione:||lug-2017|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.3390/molecules22071198|
|Appare nelle tipologie:||01 - Articolo su periodico|