title: Identification of novel scaffolds for IκB kinase beta inhibitor via a high-throughput screening TR-FRET assay.
abstract: Control of NF-κB release through the inhibition of IκB 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.
authors.affiliation: Bio-Organic Science Division, Korea Research Institute of Chemical Technology, 19 Sinseongno, Yuseong, Daejeon 305-343, Republic of Korea.
authors.forename: Kwang-Seok
authors.initials: KS
authors.lastname: Oh
authors.forename: Sunghou
authors.initials: S
authors.lastname: Lee
authors.forename: Joong Kwon
authors.initials: JK
authors.lastname: Choi
authors.forename: Byung Ho
authors.initials: BH
authors.lastname: Lee
mesh_terms: D004791:Enzyme Inhibitors / Q000032:analysis* / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D057166:High-Throughput Screening Assays; D006801:Humans; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors*
chemical_list: D004791:Enzyme Inhibitors; D051550:I-kappa B Kinase