title: TR-FRET binding assay targeting unactivated form of Bruton's tyrosine kinase.
abstract: Bruton's Tyrosine Kinase (BTK) is one of the crucial kinases for the B cell maturation and mast cell activation, and specific inhibitors of BTK are considered to be attractive targets in drug discovery research. In this Letter, we have designed and synthesized a new fluorescent probe for TR-FRET-based high-throughput screening, to identify compounds that preferentially bind to an inactive conformation of BTK which has a unique structural feature. A set of kinase-focused compound library was screened using this assay method, and compound 31 was successfully identified as a potent inhibitor which preferentially bind to the inactive conformation of BTK. These results suggest that this screening method has a great potential for the discovery of novel selective BTK inhibitors.
authors: Carna Biosciences, Inc., BMA 3F, 1-5-5 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan. Electronic address: tokiko.asami@carnabio.com.
authors: Tokiko
authors: T
authors: Asami
authors: Carna Biosciences, Inc., BMA 3F, 1-5-5 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan.
authors: Wataru
authors: W
authors: Kawahata
authors: Carna Biosciences, Inc., BMA 3F, 1-5-5 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan.
authors: Masaaki
authors: M
authors: Sawa
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D001667:Binding, Competitive; D001681:Biological Assay / Q000379:methods*; D016503:Drug Delivery Systems; D015195:Drug Design; D004789:Enzyme Activation / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry*; D020128:Inhibitory Concentration 50; D008954:Models, Biological; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*
chemical_list: D005456:Fluorescent Dyes; D047428:Protein Kinase Inhibitors; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase
keywords: Bruton’s tyrosine kinase; Fluorescent probe; Kinase inhibitor; Screening; TR-FRET