Supplementary Figure S2. NAE inhibitory effects of TAS4464. (A) E1 inhibitory effects of TAS4464 on HCT116 cells. Cells were treated with TAS4464 for 1 h. (B) Competition assay with non-labeled TAS4464. HCT116 cells were pre-treated with non-labeled TAS4464 (10, 100, 1000, 10000 nmol/L) and treated with 100 nmol/L 14C TAS4464 for 4 hours. (C) Dual-color detection of NEDD8 and UBC12. CCRF-CEM cells were treated with 100 nmol/L of TAS4464 for 1 to 24 h.
ARTICLE ABSTRACT
NEDD8-activating enzyme (NAE) is an essential E1 enzyme of the NEDD8 conjugation (neddylation) pathway, which controls cancer cell growth and survival through activation of cullin-RING ubiquitin ligase complexes (CRL). In this study, we describe the preclinical profile of a novel, highly potent, and selective NAE inhibitor, TAS4464. TAS4464 selectively inhibited NAE relative to the other E1s UAE and SAE. TAS4464 treatment inhibited cullin neddylation and subsequently induced the accumulation of CRL substrates such as CDT1, p27, and phosphorylated IκBα in human cancer cell lines. TAS4464 showed greater inhibitory effects than those of the known NAE inhibitor MLN4924 both in enzyme assay and in cells. Cytotoxicity profiling revealed that TAS4464 is highly potent with widespread antiproliferative activity not only for cancer cell lines, but also patient-derived tumor cells. TAS4464 showed prolonged target inhibition in human tumor xenograft mouse models; weekly or twice a week TAS4464 administration led to prominent antitumor activity in multiple human tumor xenograft mouse models including both hematologic and solid tumors without marked weight loss. As a conclusion, TAS4464 is the most potent and highly selective NAE inhibitor reported to date, showing superior antitumor activity with prolonged target inhibition. It is, therefore, a promising agent for the treatment of a variety of tumors including both hematologic and solid tumors. These results support the clinical evaluation of TAS4464 in hematologic and solid tumors.
