MDM2-HDAC双靶点抑制剂的设计、合成与活性研究

DESIGNESIGN, SYNTHESIS AND BIOLOGICAL EVALUATION OF DUAL – ACTING INHIBITORS AGAINAGAINST HDAC AND MDM2

黄恒军

11649070

硕士

化学

徐晶

2018

2018.7.11

哈尔滨工业大学

深圳

抑瘤蛋白p53 转录活性的激活可以诱导肿瘤细胞周期停滞和凋亡，抑制肿瘤细胞的发生与发展。MDM2 可直接与 p53 的 DNA 结合域，抑制 p53 的 转录活性，同时作为 E3 泛素连接酶诱导 p53 泛素化进而被降解。HDAC 是一类转录后修饰酶，可催化 p53 的 C-端赖氨酸残基去乙酰化，裸露的氨基一方面促进 MDM2 介导的 p53泛素化，另一方面使转录因子无法识别而抑制 p53 的转录活性。此外，HDAC还能抑制蛋白酶体介导的 MDM2 降解。因此，同时抑制MDM2 和 HDAC，则可增强 p53 在细胞核内的累积，延长 p53 的半衰期，从而实现对肿瘤细胞的持续抑制。首先，基于 HDAC 和 MDM2 两个靶点，借助计算机辅助药物设计和基于靶点的药物开发研究方法，利用 MOE 软件进行模型构建，分子模拟、对接等方法设计出 33 个目标化合物。其次，应用现代 有机 合成手段进行了目标化合物的合成，借助制备高效液相对目标化合物进行了最终纯化。最后，分别利用荧 光偏振法和 FluorFluor-de -Lys 荧光活性分析法测定了 MDM2 和 HDAC 的单点抑制活性和 IC 50。通过体外 MDM2 和 HDAC 酶活性抑制活性初步筛选得到具有潜力的目标化合物 27 -J、27 -K、27 -L和 41 ，其对 MDM2 的 IC 50 分别为 0.048 μM 、 0.070 μM 、0.064 μM 和 1.752 μM ，对 HDAC 的 IC 50为 0.508 μM 、0.321 μM 、 0. 281 μM 和 0.427 μM 。同时得出了该类双靶点抑制剂的构效关系： 1）总体来说， 母体结构为螺氧化吲哚的体外酶抑制活性强于吡咯烷衍生物； 2）连接链的 基团和长度均显著影响 HDAC 抑制活性两者在一定程度的相匹配能提高HDAC 抑制活性，不含苯环的脂肪链碳原子数为6时最佳 ；3）化合物的 HDAC 作用部位（极性区域 ）的改变对 MDM2 抑制活性影响不大，但母体结构的不同对其影响显著。本课题首次进行了MDM2 和 HDAC 双靶点抑制剂的探索，获得了可进一步开展细胞实验的三个候选化合物，为开发出疗效高、副作用小的抗癌候选药提供了新的思路，具有明显的现实意义与社会意义！

The tumor suppressor protein p53 plays a pivotal role in the regulation of cellular process and suppression of tumor development. As a transcriptional factor, p53 is activated under cellular stress and regu lates multiple downstream genes implicated in cell cycle control, apoptosis, DNA repair, and senescence. While p53 is inactivated in about 50% of human malignancies by mutation or deletion, it remains wild wild-type in the other half cases but its function is i mpaired by other mechanisms. One such mechanism is the over expression of MDM2, the primary negative regulator of p53, which effectively inhibits p53 function. As an E3 ubiquitin ligase, MDM2 can directly interrupt p53 transcriptional activity through bind ing to the N N-terminal transcription activation domain of p53 and subsequently inducing ubiquitin ubiquitin-dependent degradation of p53. It was reported that acetylated p53 lysine overlap with those ubiquitylated, and MDM2 can promote p53 degradation by recruiting H DAC complex to remove these acetyl groups. In addition, HDAC can inhibit proteasome proteasome-mediated degradation of MDM2. Thus, a small molecule targeting MDM2 and HDAC simultaneously might exert a combination effect in the cancer treatment via increasing p53 prot ein level in cell nucleus and extending its halfhalf-life.Based on the aid of computer aided drug design methods, 3 33 compounds targeting MDM2 and HDAC were designed. And those compounds were synthesiszed and biological evaulated through modern synthetic metho ds and testing enzyme activity inhibition rate, respectively. Among 33 compounds , compounds 27 -J, 27 -K, 27 -L and 41 were found to inhibit MDM2 with IC IC50 of 0.048 μM , 0. 070 μM , 0.0 64 μM and 1.752 μM and HDAC with IC IC50 of 0.508 μM , 0.321 μM , 0. 281 μM and 0.427 μM , respectively. Preliminary structure structure-activity relationships studies showed that: (1) compounds bearing spiro spiro-oxindoles were more active than pyrrolidone derivatives; (2) compounds bearing aliphatic chains with six carbons showed better HDAC inhibi tory activities; (3) the difference of parent structures had significant influence on MDM2 inhibitory activities.A thorough survey of literature indicated that there was no reported research that was oriented to target MDM2 MDM2-HDAC. It provides a new way of thinking to develop anticancer candidate drugs with high curative effect and small side effect, which has obvious practical and social significance.

p53 双靶点抑制剂 HDAC MDM2 构效关系

p53 Dual target inhibitorsinhibitors HDAC MDM2 QSAR

中文

联合培养

南方科技大学

黄恒军. MDM2-HDAC双靶点抑制剂的设计、合成与活性研究[D]. 深圳. 哈尔滨工业大学,2018.