第二代RhoA小分子抑制剂的发现及其抗血管痉挛活性研究

发布时间：2018-03-26 05:13

  本文选题：脑血管痉挛　切入点：蛛网膜下腔出血　出处：《华东理工大学》2014年硕士论文

【摘要】：血管痉挛是心脑血管疾病重要的病理生理机制之一,开发针对新靶标和具有新作用机制的解除血管痉挛的治疗药物具有重要的科学意义。RhoA/ROCK系统是细胞收缩不依赖于细胞内钙浓度变化的重要信号传导途径,该通路相关蛋白的高表达或过度激活与血管痉挛的发生有着重要的关系,因此RhoA蛋白可能是一个潜在的药物作用新靶点。考虑到目前国际国内有关直接针对RhoA蛋白GTP结合域的抑制剂研究报道很少的状况,在课题组前期发现的第一代RhoA小分子抑制剂结构和药效团信息基础上,本论文开展了第二代RhoA小分子抑制剂的设计、合成、体外内药理研究。基于应用分子对接技术模拟出的RhoA与第一代抑制剂的相互作用特征,本论文确定了第二代小分子RhoA抑制剂的设计思路,共设计合成34个目标物(A01-A34)。对所有34个目标物开展了细胞水平的RhoA抑制活性研究,成功发现了6个目标物(A01、A02、A07、A12、 A14)为强效RhoA抑制剂,它们的IC50值处于1～2μM之间,可以定义为第二代RhoA小分子抑制剂。进一步测试了6个RhoA抑制剂对血管张力的影响,发现2个目标物对苯肾上腺素预制血管收缩有明显的舒张效果(A02:IC50=70.9±1.3μM; A14: IC50=73.4±2.6κM),其舒张血管效果相比第一代RhoA的抑制提高了大约一倍左右。通过考察A02、A10和A14这3个目标物在水中的溶解性,我们最终选择水溶性良好(S=639μg/mL)的目标物A02开展初步的体内药效研究。在蛛网膜下腔出血后脑血管痉挛大鼠模型中,第二代RhoA抑制剂A02对SD大鼠SAH-CVS有明显的缓解作用,在低剂量组(95mM)表现出与临床药物法舒地尔(15.2mM)相当的药理效果。根据细胞、组织和动物三种水平药理活性结果,我们总结归纳了本论文第二代RhoA抑制剂的构效关系。这些SARs结果为今后寻找小分子RhoA抑制剂提供了结构线索。同时,本论文发现的强效第二代RhoA小分子抑制剂A02可以作为具有自主知识产权的新型脑血管痉挛治疗临床前候选药物进行下一步的开发研究。
[Abstract]:Vasospasm is one of the important pathophysiological mechanisms of cardiovascular and cerebrovascular diseases. It is of great scientific significance to develop new targets and new mechanisms for the treatment of vasospasm. RhoA / rock system is an important signal transduction pathway for cell contraction independent of changes in intracellular calcium concentration. The overexpression or overexpression of proteins associated with this pathway is associated with the development of vasospasm. Therefore, RhoA protein may be a potential new target for drug action. Considering that there are few reports on inhibitors directly targeting the GTP binding domain of RhoA protein at home and abroad, Based on the information of the structure and pharmacophore of the first generation of RhoA small molecule inhibitors, the design and synthesis of the second generation of RhoA small molecule inhibitors were carried out in this paper. In vitro pharmacological studies. Based on the interaction characteristics of RhoA and first-generation inhibitors simulated by molecular docking technique, the design idea of the second generation of small molecular RhoA inhibitors was determined in this paper. A total of 34 target compounds (A01-A34) were designed and synthesized. Cell-level RhoA inhibitory activity of all 34 targets was studied. It was successfully found that 6 target compounds (A01A02A02A07A12, A14) were potent RhoA inhibitors, and their IC50 values were in the range of 1 渭 M. It can be defined as the second generation of RhoA small molecule inhibitors. The effects of 6 RhoA inhibitors on vascular tension were further tested. It was found that the vasodilation effects of the two target compounds on the prefabricated vasoconstriction of phenylephrine were 70.9 卤1.3 渭 M and A14: IC50=73.4 卤2.6 渭 M, respectively. The inhibition of the vasodilation effect of the two target compounds was about twice as high as that of the first generation of RhoA. The solubility of A02: A10 and A14 in water was investigated. We finally selected A02, a water-soluble target of 639 渭 g 路mL ~ (-1), to study its pharmacodynamics in vivo. In the model of cerebral vasospasm after subarachnoid hemorrhage, A02, a second-generation inhibitor of RhoA, could significantly relieve SAH-CVS in SD rats. In the low dose group (95 mm), the pharmacological effect was comparable to that of the clinical drug fasudil 15.2mM.According to the results of three levels of pharmacological activity of cells, tissues and animals, We summarize the structure-activity relationships of the second generation of RhoA inhibitors in this thesis. These SARs results provide a structural clue for finding small molecular RhoA inhibitors in the future. A02, a potent second generation small molecule inhibitor of RhoA, can be used as a new pre-clinical candidate for cerebral vasospasm therapy with intellectual property rights.
【学位授予单位】：华东理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

【参考文献】

相关博士学位论文 前1条

1 邓菁;基于结构的药物设计和优化及两种有机小分子不对称催化反应研究[D];华东理工大学;2012年

，

本文编号：1666483