Characterizing Small Molecule Inhibitors of ALDH1A1 with Biochemical, Biophysical, and High Throughput Cell-based Assays

Authors

• AS Yasgar
• SM Yang
• Y Wang
• C Danchik
• SA Titus
• B Baljinnyam
• DM Cheff
• XS Wang
• DJ Maloney
• A Jadhav
• M Lal-Nag
• V Vasilou
• U Opperman
• A Simeonov
• NJ Martinez

Abstract

Aldehyde dehydrogenases (ALDHs) have a broad spectrum of biological activities through the oxidation of both endogenous and exogenous aldehydes and possess important physiological and toxicological functions in areas such as CNS, metabolic disorders, and cancers. Increased expression of ALDH1A1 has been identified in a wide-range of human cancer stem cells and is associated with cancer relapse and poor prognosis, highlighting the need for the identification and development of small molecule ALDH inhibitors. To facilitate quantitative high-throughput screening (qHTS) campaigns for the discovery, characterization and structure-activity-relationship (SAR) studies of small molecule ALDH1A1 inhibitors, we show the miniaturization to 1,536-well format of an enzymatic and high-content cell-based ALDEFLUOR assay. We demonstrate the utility of these assays by generating dose-response curves on a comprehensive set of prior art inhibitors and in-house synthesized analogs. Finally, to determine residence times of lead compounds, we further characterized compounds using Surface Plasmon Resonance (SPR), to determine binding to ALDH1A1 with the aim of comparing residence times

Scientific Focus Area: Chemical Biology

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