title: [3H]BMS-599240--a novel tritiated ligand for the characterization of BACE1 inhibitors.
abstract: In this report we describe a novel radioligand, [(3)H](S)-2-((S)-3-Acetylamino-3-sec-butyl-2-oxo-pyrrolidin-1-yl)-N-[(1S,2R)-1-benzyl-2-hydroxy-3-(3-methoxy-benzylamino)-propyl]-4-phenyl-butyramide ([(3)H]BMS-599240), that exhibits robust specific binding in homogenates from cell cultures overexpressing beta-site amyloid precursor protein cleaving enzyme-1 (BACE1). Radioligand binding exhibited high affinity, K(d)=2 nM, commensurate with its inhibitory potency against BACE1. Inhibition of radioligand binding in the presence of a range of different BACE1 inhibitors exhibited the same rank order of potency as for inhibition of BACE1 enzymatic activity. BACE1-dependent binding of the radioligand was also demonstrated in mouse brain homogenates, where genetic ablation of BACE1 eliminated high affinity binding. Thus, the radioligand [(3)H]BMS-599240 is a novel tool potentially useful for evaluation of BACE1 enzyme in biological samples, and for evaluation of inhibitor binding to BACE1.
authors.affiliation: Neuroscience Biology, Bristol-Myers Squibb Research and Development, Wallingford, CT 06492, USA.
authors.forename: Lawrence G
authors.initials: LG
authors.lastname: Iben
authors.forename: Lisa
authors.initials: L
authors.lastname: Kopcho
authors.forename: Jovita
authors.initials: J
authors.lastname: Marcinkeviciene
authors.forename: Changsheng
authors.initials: C
authors.lastname: Zheng
authors.forename: Lorin A
authors.initials: LA
authors.lastname: Thompson
authors.forename: Charles F
authors.initials: CF
authors.lastname: Albright
authors.forename: Jeremy H
authors.initials: JH
authors.lastname: Toyn
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008024:Ligands; D051379:Mice; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D011760:Pyrrolidinones; D011869:Radioligand Assay
chemical_list: C532840:((3)H)(S)-2-((S)-3-acetylamino-3-sec-butyl-2-oxo-pyrrolidin-1-yl)-N-((1S,2R)-1-benzyl-2-hydroxy-3-(3-methoxy-benzylamino)-propyl)-4-phenyl-butyramide; D018076:DNA, Complementary; D008024:Ligands; D016271:Proto-Oncogene Proteins c-myc; D011760:Pyrrolidinones; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; C509284:BACE2 protein, human; C509280:BACE1 protein, human