Nutlin-3a is the active enantiomer of Nutlin-3, an MDM2 antagonist that inhibits MDM2-p53 interaction (Ki=90 nM) and activates p53. Nutlin-3a binds preferentially to the p53-binding pocket of MDM2, leading to stabilization of p53 and activation of the p53 pathway. Nutlin-3a has antitumor activity.

MDM2-p53 interaction:90 nM

方法：人胶质母细胞瘤细胞 U87MG (p53 wild-type) 和 T98G (p53-mutant) 用 Nutlin-3a (0.6-20 µM) 处理 96 h，通过 MTT assay 检测细胞活力。
结果：与 Nutlin-3a 孵育 96 h 导致野生型 p53 U87MG 细胞系中细胞存活率呈剂量依赖性降低，而突变型 p53 T98G 细胞系中没有观察到细胞存活率的显著降低。[1]
方法：U-2 OS 细胞用 Nutlin-3a (2-10 µM) 处理 48 h，通过 flow cytometry 检测细胞凋亡和细胞周期。
结果：在 U-2 OS 细胞中检测到约 37% 的凋亡细胞。我们还发现，Nutlin-3a 处理的 U-2 OS 细胞以剂量依赖的方式阻滞在 G1 期。[2]

方法：为检测体内抗肿瘤活性，将 Nutlin-3a (25 mg/kg) 腹腔注射给携带 U-2 OS (wt p53) 异种移植物的 SCID 小鼠，每天一次，持续两周。
结果：治疗 2 周后，对于 U-2 OS 细胞，Nutlin-3a 治疗组的平均肿瘤体积为 480 mm3，而对照组为 1751 mm3。此外，两只接受 Nutlin-3a 治疗的小鼠在治疗结束时没有明显的肿瘤。[2]

Biacore studies: Competition assays are performed on a Biacore S51. A Series S Sensor chip CM5 is derivatized for immobilization of a PentaHis antibody for capture of the His-tagged p53. The level of capture is ~ 200 response units (1 response unit corresponds to 1 pg of protein per mm 2). The concentration of MDM2 protein is kept constant at 300 nM. Test compounds are dissolved in DMSO at 10 mM and further diluted to make a concentration series of inhibitor in each MDM2 test sample. The assays are run at 25 °C in running buffer (10 mM Hepes, 0.15 M NaCl, 2% DMSO). MDM2-p53 binding in the presence of inhibitor is calculated as a percentage of binding in the absence of inhibitor and IC50 is calculated using Microsoft Excel

SRB(Only for Reference)