Discovery and Characterization of Novel Nonsubstrate and Substrate NAMPT Inhibitors

Abstract

Cancer cells are highly reliant on NAD⁺-dependent processes, including glucose metabolism, calcium signaling, DNA repair, and regulation of gene expression. Nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme for NAD⁺ salvage from nicotinamide, has been investigated as a target for anticancer therapy. Known NAMPT inhibitors with potent cell activity are composed of a nitrogen-containing aromatic group, which is phosphoribosylated by the enzyme. Here, we identified two novel types of NAM-competitive NAMPT inhibitors, only one of which contains a modifiable, aromatic nitrogen that could be a phosphoribosyl acceptor. Both types of compound effectively deplete cellular NAD⁺, and subsequently ATP, and produce cell death when NAMPT is inhibited in cultured cells for more than 48 hours. Careful characterization of the kinetics of NAMPT inhibition <i>in vivo</i> allowed us to optimize dosing to produce sufficient NAD⁺ depletion over time that resulted in efficacy in an HCT116 xenograft model. Our data demonstrate that direct phosphoribosylation of competitive inhibitors by the NAMPT enzyme is not required for potent <i>in vitro</i> cellular activity or <i>in vivo</i> antitumor efficacy. <i>Mol Cancer Ther; 16(7); 1236-45. ©2017 AACR</i>.