Title:Discovery of Novel Scaffolds for Rho Kinase 2 Inhibitor Through TRFRET- Based High Throughput Screening Assay
Volume: 16
Issue: 1
Author(s): Kwang-Seok Oh, Jihye Mun, Jae Eun Cho, Sunghou Lee, Kyu Yang Yi, Chae Jo Lim, Jin Soo Lee, Whui Jung Park and Byung Ho Lee
Affiliation:
Keywords:
High throughput screening, Immobilized metal affinity for phosphochemicals, rho kinase, ROCK, ROCK2
inhibitor, time-resolved fluorescence resonance energy transfer, IMAP-TR-FRET assay, Dimethyl sulfoxide, Dithiothreitol, Protein kinase C, Serum and glucocorticoid-induced protein kinase.
Abstract: Recent advances in basic and clinical studies have identified Rho kinase (ROCK) as an important target
potentially implicated in a variety of cardiovascular diseases and ROCK inhibitors were considered as a pharmacological
strategy to prevent and treat cardiovascular diseases. To screen the small molecule compound library against ROCK, a
high throughput screening (HTS) campaign was carried out using immobilized metal affinity for phosphochemicals
(IMAP)-based time-resolved fluorescence resonance energy transfer (TR-FRET) assay. Z’ value and signal to background
(S/B) ratio were achieved at 0.76 and 5.27 for the pilot library screening of the most diverse set consisting of 15,040
compounds with a reasonable reconfirmation rate. From this screening campaign, four novel scaffolds, such as 3-
nitropyridine, 4-methoxy-1,3,5,-triazine, naphthalene-1,4-dione, and 2,3-dihydro-1H-pyrrolo[2,3-b]quinoxaline, were
yielded. Particularly, we found that 3-nitropyridine derivatives possess potent inhibitory activity and selectivity for
ROCK. Our findings provide important information for the design of novel ROCK inhibitor.