Pevonedistat (MLN4924) is a potent and selective NEDD8 activating enzyme (NAE) inhibitor (IC50=4.7 nM). Pevonedistat can be used for the treatment of myelodysplastic syndromes (MDS) and also has antitumor activity.

CAPAN-1 cells:101 nM, BEAS-2B cells:1.22 μM, HGC-27 cells:1.25 μM, GES1 cells:13.45 μM, Neuroblastoma:136-400 nM, NAE:4.7 nM, HepG2 cells:0.3 μM, EKVX cells:0.3 μM, HuTu80 cells:42.6 nM, Huh-7 cells:0.3 μM, HCT116 cells:33.89 nM, Caco-2 cells:4.4 μM, HT29 cells:484 nM, A549 cells:0.63 μM, DU-145 cells:201 nM

Bcl-2 Binding affinity calculation: A predicted binding affinity for Obatoclax binding to BCL-2 is calculated using the SIE scoring function. [4] As a control in determining the reliability of the calculation, predicted binding affinities Ki) are calculated for a set of 12 small molecules with experimentally measured binding affinities to BCL-2.

MLN4924 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[1]. HCT-116 cells grown in 6-well cell-culture dishes are treated with 0.1% DMSO (control) or 0.3 μM MLN4924 for 24 h. Whole cell extracts are prepared and analysed by immunoblotting. For analysis of the E2-UBL thioester levels, lysates are fractionated by non-reducing SDS-PAGE and immunoblotted with polyclonal antibodies to Ubc12, Ubc9 and Ubc10. For analysis of other proteins, lysates are fractionated by reducing SDS-PAGE and probed with primary antibodies as follows: mouse monoclonal antibodies to CDT1, p27, geminin, ubiquitin, securin/PTTG and p53 or rabbit polyclonal antibodies to NRF2, Cyclin B1 and GADD34[1].