Title: Identification of Novel Scaffolds for IκB Kinase Beta Inhibitor via a High Throughput Screening TR-FRET Assay
VOLUME: 13 ISSUE: 9
Author(s):Kwang-Seok Oh, Sunghou Lee, Joong Kwon Choi and Byung Ho Lee
Affiliation:Bio-Organic Science Division, Korea Research Institute of Chemical Technology, 19 Sinseongno, Yuseong, Daejeon, 305-343, Republic of Korea.
Keywords:IκB kinase β, IKKβ inhibitor, time-resolved fluorescence resonance energy transfer.
Abstract: Control of NF-κB release through the inhibition of I & κB kinase β (IKKβ) has been identified as a potential target for the treatment of inflammatory and autoimmune diseases. To screen the small molecule compound library against IKKβ, 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. Through serial optimization of assay conditions, the Z value was achieved at 0.88 from the pilot library screening of the most diverse 7,243 compounds with reconfirmation rate of 63%. The results from this HTS campaign identified three novel scaffolds for the prospective IKKβ inhibitor, such as 7-benzoyl-4-phenylcyclopenta [1,2] oxazine, 1-(thiophen or furan)-2,3- dihydroimidazo[1,5] pyridine and 2-phenyloxazolo [5,4] pyridine. Particularly, 7-benzoyl-4-phenylcyclopenta [1,2] oxazine derivatives presented potent inhibitory activity and selectivity for IKKβ. These findings suggest that the current TR-FRET assay system for IKKβ was successful to identify hits for novel IKKβ inhibitors as a robust, reproducible and sensitive HTS system.
