Caracterización de inhibidores de la actividad de canales iónicos sensibles a voltaje: sensores de voltaje (fluorescentes) vs. electrofisiología

Abstract

The aim of the development of these investigation’s projects in Almirall is to find new molecules for a particular disease and it can get started in two different ways: the selection of a phenotype or of a protein targeting (GPCRs, enzymes, ion channels, etc.) whose modulation could become a benefit to a pathology of interest. One of the proteins targeting that is currently raising more interest in the therapeutic investigation are the ion channels. The reason is that they play a critical role in the monitoring of many physiological processes. These processes form the molecular base of the nervous system, they also adjust the selective transport of the ions and macromolecules, they have a vital role in the synthesis of the mediators, they take part in the homeostasis and in the cellular communication. This project focus in the (VGSC). Nine sodium channels in humans have been identified and found nowadays that perform vital functions in the central nervous system and in the peripheral nervous system, as well. The dysfunction of these channels can create diseases which are called channelopathies such as the Brugada syndrome, childhood epilepsy or chronic pain. Due to the required need in the discover of new drugs, the main objective of the study that was carried out for Almirall focused in the set-up of a new primary high-efficiency assay and minor costs than the electrophysiology using the platform FLIPR Tetra and monitoring the changes in the membrane potential, that could allow to characterize the power of new inhibitors of the sodium channel Nav1x. Once the set-up of the FLIPR Tetra was done, twenty-four inhibitors that were previously tested with the electrophysiology IonWorks quatro were characterized. The results obtained with the FLIPR Tetra were in truth successful, the 87,5% being the most detected compounds in the same potency order than the electrophysiology. Moreover, the 75% of the detected compounds with a different potency order were more powerful than in the electrophysiology assay and the remaining 25% only seven times less powerful. This could assure its detection with the set-up assay. The controlling assay of the membrane potential using the FLIPR Tetra could be considered acceptable as a primary assay of characterization and narrowed down for specific inhibitors of the sodium channel Nav1.x, putting the electrophysiology as a activityconfirmation assay and characterization of inhibitors