【作者】 陈明；

【导师】 姜凤超；

【作者基本信息】 华中科技大学 ， 药物化学， 2016， 硕士

【摘要】 整合素是细胞表面的一种重要跨膜受体,通过介导细胞与细胞外基质及细胞与细胞间的双向信号传导,在肿瘤细胞迁移、粘附、浸润及肿瘤血管新生中起重要作用。将整合素αvβ3作为靶点进行的抗肿瘤药物的研发及相对应治疗方法的开发已成为当前肿瘤治疗的热点之一。利用计算机辅助药物设计软件Catalyst选择实验中与αvβ3受体易于结合且有较高抑制活性30个化合物作为训练集,构建整合素αvβ3受体抑制剂最佳药效团模型。通过已知的整合素αvβ3内源性配体部位RGD片段为先导化合物,结合所设计的药效团模型进行结构优化,最后通过结构改造得到16个未见文献报道的目标化合物过常规方法和微波辐射方法合成得到目标化合物,利用UV、IR、1H-NMR、13C-NMR、ESI-MS分析手段对目标化合物进行结构确证并通过HPLC确定目标产物的纯度。在肿瘤新生血管生成的过程中考虑到αvβ3受体在增殖的血管内皮细胞上表达比较高,所以本实验选用肿瘤血管内皮细胞相当的人脐静脉内皮细胞(HUVEC)作为药理模型对目标产物进行初步活性筛选。为了进一步验证目标化合物的抗肿瘤作用,本论文还选用了αvβ3表达比较高的其他两株肿瘤细胞胰腺癌细胞PANC-1,黑色素瘤细胞A375,采用MTT方法检测目标化合物的药理活性。实验结果显示大部分目标化合物对高度表达αvβ3受体的HUVEC细胞和较高表达αvβ3两株肿瘤细胞A375和PANC-1的都有一定的抑制作用,对HUVEC细胞有三个化合物活性高于阳性对照,对A375有两个化合物的活性高于阳性对照,其中活性最高的化合物E2对HUVEC和A375的IC50值分别为0.023μmol/L,0.41μmol/L。综上所述,本研究的创新之处在于:构建整合素αvβ3受体拮抗剂的最佳药效团模型,设计并合成了16个对整合素αvβ3受体具有较强拮抗作用的苯基哌嗪类衍生物,通过药理实验表明对肿瘤细胞有一定的抑制作用,为αvβ3受体拮抗剂小分子活性药物的设计和改造提供一定的理论和实验依据。更多还原

【Abstract】 Integrins are important adhesion molecules located on cell surface,which can mediate cell to cell and cell to extracellular matrix(ECM) interaction through bidirectional signal transduction and plays an important role in tumor ce lls migration, adhesion, invasion and angiogenesis. Integrin αvβ3 overexpressed in many tumor cell surface while low or no expression in normal cells, when its function is inhibited, vascular endothelial cell apoptosis, tumor growth was inhibited, and e ven tumor regression. The research using integrins αvβ3 as targets on anti- tumor drug design and the corresponding treatment strategy has been a hot point in recent years.In our study, a pharmacophore model of integrin αvβ3 receptor inhibitors were constructed based on currently available 30 inhibitors with highly inhibitory activity against integrin αvβ3 receptor in Catalyst software. The tripeptide RGD sequence were chosen as a scaffold for further chemical modification by phamacophore model to explore αvβ3 inhibitors. Finally,sixteen novel target compounds have been designed and synthesized after further modification of the structure of the leading compounds and sixteen were whole new and not reported. 16 target compounds were synthesized by conventional methods or microwave-catalized organic synthesis method. UV、IR、1H-NMR、13C-NMR 、ESI-MS、 HPLC were applied to confirm the structure and purity of target compounds. Integrin αvβ3 overexpressed in tumor angiogenesis process, so this experiment, HUVEC as useful primary indicator. To further verify the antitumor effect of the target compound, the paper also selected αvβ3 expression higher tumor cells PANC-1, A375, the inhibitory activity to three cells of target compounds was evaluated by MTT. The results of antitumor activities show that the majority of the target compounds have good inhibitory activity, some of the compounds have higher activity than the positive control. Compound E2 has the maximum inhibition of A375 and HUVEC with the IC50 value of 0.41 μmol/L, 0.023 μmol/L.In conclusion, the innovate points of this research was to have built an optimum integrin αvβ3 receptor antagonist pharmaphore model, design and synthesis sixteen unreported compounds of phenylpiperazine derivatives which have better antagonism forintegrin αvβ3 receptor. Pharmacology experiments indicated that those compounds have certain inhibition for some cancer cell growth. The results have provide a theoretical and experimental basis of small molecule antagonists of the αvβ3 active drug design and renovation.更多还原