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.

NAE:4.7 nM

方法：9 种神经母细胞瘤细胞系用 Pevonedistat (12-1000 nM) 处理 72 h，使用 MTT 方法检测细胞活力。
结果：所有测试的神经母细胞瘤细胞系对 Pevonedistat 敏感，IC50 值范围为 136-400 nM。[1]
方法：人结直肠癌细胞 HCT-116 用 Pevonedistat (0.001-3 μM) 处理 24 h，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：Pevonedistat 处理导致 Ubc12–NEDD8 硫酯和 NEDD8-cullin 偶联物的剂量依赖性降低，导致已知 CRL 底物 CDT1、p27 和 NRF2 的丰度增加。[2]

方法：为检测体内抗肿瘤活性，将 Pevonedistat (30-60 mg/kg in 10% cyclodextrin) 皮下注射给携带人结直肠癌肿瘤 HCT-116 的 BALB/c 小鼠，每天一次或两次，持续二十天。
结果：Pevonedistat 抑制 HCT-116 异种移植瘤生长。[2]
方法：为检测体内抗肿瘤活性，将 Pevonedistat (60 mg/kg，腹腔注射，每天一次)、venetoclax (50 mg/kg，口服给药，每天一次) 和 azacitidine (8 mg/kg，静脉注射，每七天三次) 给药给携带 AML 肿瘤 OCI-AML2-Red-Fluc 的 NSGS 小鼠，持续十四天。
结果：Pevonedistat/venetoclax/azacitidine 三联治疗在急性髓系白血病的全身异种移植物模型中诱导持久反应。[3]

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].