[3H]BMS-599240 — A novel tritiated ligand for the characterization of BACE1 inhibitors

In this report we describe a novel radioligand, [3H](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 ([3H]BMS-599240), that exhibits robust specific binding in homogenates from cell cultures overexpressing β-site amyloid precursor protein cleaving enzyme-1 (BACE1). Radioligand binding exhibited high affinity, Kd = 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 [3H]BMS-599240 is a novel tool potentially useful for evaluation of BACE1 enzyme in biological samples, and for evaluation of inhibitor binding to BACE1.