Design, synthesis and biological evaluation of a novel tubulin inhibitor 7a3 targeting the colchicine binding site
Design, synthesis and biological evaluation of a novel tubulin inhibitor 7a3 targeting the colchicine binding site
复制标题
一种针对秋水仙碱结合位点的新型微管蛋白抑制剂 7a3 的设计、合成和生物学评价
DOI：
10.1016/j.ejmech.2018.05.010
复制
发表时间：
2018-08-05
影响因子：
6.7
通讯作者：
Yang, Jinliang
中科院分区：
文献类型：
--
作者：
Lai, Qinhuai;Wang, Yuxi;Yang, Jinliang
靶向秋水仙碱结合位点的微管蛋白抑制剂继续成为有前途的抗癌剂。本研究以考布他汀A-4的顺式吡唑类似物为基础，设计、合成并优化了一个以秋水仙碱结合位点为靶点的新型微管蛋白抑制剂7a 3。这些新合成的化合物的构效关系（SAR）进行了总结，表明在N1位的甲基取代基和脱氨基是显着重要的抗增殖功效。优化的化合物7a 3显示出将细胞周期阻滞在G2/M期、诱导细胞凋亡和抑制肿瘤细胞的细胞迁移的能力。共聚焦显微镜免疫荧光分析和微管蛋白聚合试验结果表明，微管蛋白组装在体外被7a 3破坏。通过解析微管蛋白复合物7a 3的晶体结构（PDB code 5 Z4 U），进一步验证了分子对接的结果，并进一步证明了7a 3与秋水仙碱的结合位点。此外，小鼠血浆中的药代动力学分析显示，7a 3在腹膜内给药后0.25 h迅速达到峰浓度，T-1/2、C-max和AUC（0-inf）分别为1.67 +/- 0.28 h、882 +/- 71 ng mL（-1）和1166 +/- 129 h ng.mL（-1），单次给药后，通过液相色谱-串联质谱法（LC/MS/MS）进行分析。此外，体内研究表明7a 3显著抑制裸鼠模型中SK-0 V-3异种移植物的肿瘤生长。总之，我们的研究证明7a 3是一个潜在的微管靶向药物用于癌症治疗。基于X射线共晶结构的7a 3的SAR和作用机制研究为下一代微管蛋白抑制剂用于癌症治疗提供了见解。(C)2018 Elsevier Masson SAS。All rights reserved.
Tubulin inhibitors that target the colchicine binding site continue to emerge as promising anticancer agents. In this study, based on the anti-proliferative activities, a novel tubulin inhibitor 7a3 targeting the colchicine binding site was designed, synthesized, and optimized from a series of novel cis-restricted pyrazole analogues of combretastatin A-4. The structure-activity relationships (SARs) of these newly synthesized compounds are summarized indicating that the methyl substituent at the N1 position and deamination were significantly important for the anti-proliferative efficacy. The optimized compound 7a3 exhibited the ability to arrest the cell cycle in the G2/M phase, induce cell apoptosis, and inhibit cell migration in tumour cells. The results of the immunofluorescence analysis using confocal microscopy and the tubulin polymerization assay revealed that tubulin assembly was disrupted by 7a3 in vitro. Furthermore, the targeting identification of 7a3 was illuminated by solving the crystal structure of 7a3 in complex with tubulin at a resolution of 3.2 angstrom (PDB code 5Z4U), which confirmed the result of molecular docking and further demonstrated that 7a3 binds to the site of colchicine. Moreover, the pharmacokinetic analysis in mouse plasma showed that 7a3 rapidly reached a peak concentration at 0.25 h after intraperitoneal administration, and the T-1/2, C-max, and AUC(0-inf), were 1.67 +/- 0.28 h, 882 +/- 71 ng mL(-1), and 1166 +/- 129 h ng.mL(-1), respectively, after a single-dose administration analysed by liquid chromatography-tandem mass spectrometry (LC/MS/MS). In addition, the in vivo study indicated that 7a3 significantly inhibited the tumour growth of the SK-OV-3 xenograft in a nude mouse model. In conclusion, our study proved 7a3 to be a potential microtubule-targeting drug for cancer therapy. The SARs and mechanism of action studies of 7a3 based on the X-ray co-crystal structure provided insights into the next-generation tubulin inhibitors for cancer therapy. (C) 2018 Elsevier Masson SAS. All rights reserved.