|Publication Type||Journal Article|
|Year of Publication||2006|
|Authors||Brandish, PE, Chiu, C-S, Schneeweis, J, Brandon, NJ, Leech, CL, Kornienko, O, Scolnick, EM, Strulovici, B, Zheng, W|
|Journal||J Biomol Screen|
|Date Published||2006 Aug|
|Keywords||Animals, Automation, CHO Cells, Cricetinae, D-Amino-Acid Oxidase, Drug Evaluation, Preclinical, Enzyme Inhibitors, Humans, Models, Biological, Tissue Array Analysis, Transfection|
Enzymes are often considered less "druggable" targets than ligand-regulated proteins such as G-protein-coupled receptors, ion channels, or other hormone receptors. Reasons for this include cellular location (intracellular vs. cell surface), typically lower affinities for the binding of small molecules compared to ligand-specific receptors, and binding (catalytic) sites that are often charged or highly polar. A practical drawback to the discovery of compounds targeting enzymes is that screening of compound libraries is typically carried out in cell-free activity assays using purified protein in an inherently artificial environment. Cell-based assays, although often arduous to design for enzyme targets, are the preferred discovery tool for the screening of large compound libraries. The authors have recently described a novel cell-based approach to screening for inhibitors of a phosphatase enzyme and now report on the development and implementation of a homogeneous 3456-well plate assay for D-amino acid oxidase (DAO). Human DAO was stably expressed in Chinese hamster ovary (CHO) cells, and its activity was measured as the amount of hydrogen peroxide detected in the growth medium following feeding the cells with D-serine. In less than 12 weeks, the authors proved the concept in 96-and then 384-well formats, miniaturized the assay to the 3456-well (nanoplate) scale, and screened a library containing more than 1 million compounds. They have identified several cell-permeable inhibitors of DAO from this cell-based high-throughput screening, which provided the discovery program with a few novel and attractive lead structures.
|Alternate Journal||J Biomol Screen|