We will be participating in SLAS2016 Conference (San Diego, Jan 23-27, 2016), with a poster “Discovery of Novel SIRT1 Modulators Using Differential Scanning Fluorimetry” (№2125). Our poster presentation is scheduled on January 25, 1-3 р.m.
Discovery of Novel SIRT1 Modulators Using Differential Scanning Fluorimetry
Sergey Zozulya, Ph.D.
78 Chervonotkatska Str., Kyiv 02094, Ukraine
Co-Authors: Roman Stavniichuk, Petro Borysko, Anastasia Griniukova, Maria Kliachina, Olexander Vasylchenko, Olexii Balinskyi, Ihor Zahanich, Alex Kiselyov
Sirtuins (SIRTs), a subfamily of histone deacetylases (HDACs), are key therapeutic targets involved in metabolic diseases, neurodegeneration, cancer, inflammation and regulation of aging and lifespan. Both inhibitors and activators of SIRT including modulators of relevant protein-proteins interactions (ex., DBC1, AROS) are of interest. The discovery of novel active compounds exemplified by SIRT ‘activators’ in particular, is hampered by the intrinsic limitations and issues associated with conventional biochemical assays. Here, we report the use of a label-free, biophysical binding assay (Differential Scanning Fluorimetry, Thermal Shift Assay) for primary screening of diverse, lead-like library of 9,000 small molecules against recombinant human SIRT1. This set was selected from Enamine’s 2,000,000 compound collection using proprietary i) structure-based, ii) ligand-based in silico discovery tools, as well as a series of medicinal chemistry filters. Screening yielded ca. 50 chemically diverse TSA hits displaying thermal shifts of 0.5-2°C and representing 4 distinct chemical classes. These SIRT1 binders were further characterized via the combination of a conventional enzymatic biochemical SIRT1 assay, LC-MS based peptide substrate conversion assay and cell-based assays to arrive at validated optimization candidates. In this communication, we provide preliminary insight into the SIRT1 modulatory effects displayed by these compounds and their tentative mechanisms of action.