本文選題：hERG鉀離子通道 + 小分子熒光探針　； 參考：《山東大學(xué)》2016年碩士論文

【摘要】：hERG (human ether-a-go-go related gene)編碼的快速激活型延遲整流鉀電流(IKr)鉀離子通道在心臟動(dòng)作電位復(fù)極化過程中具有關(guān)鍵作用。當(dāng)hERG發(fā)生突變或hERG通道被藥物阻滯時(shí),可引起遺傳性或獲得性長QT間期綜合征(LQTS)。由于不同種類藥物對hERG通道產(chǎn)生阻滯作用而引起長QT綜合征,藥物的安全性已經(jīng)在藥物研發(fā)和臨床研究中引起關(guān)注,因此hER G通道已經(jīng)作為候選藥物心臟毒性篩選的重要靶點(diǎn)。此外,初步研究表明hERG在某些腫瘤細(xì)胞中的表達(dá)顯著上調(diào),而在腫瘤細(xì)胞相應(yīng)的正常細(xì)胞中不表達(dá)或表達(dá)較低,而且發(fā)現(xiàn)hERG通道與腫瘤的分化、凋亡、繁殖等密切相關(guān),預(yù)示hERG將有可能作為腫瘤的新型生物標(biāo)記物。因此,對hERG通道的系統(tǒng)生理病理研究將對新藥臨床前研究中的心臟毒性安全性評價(jià)和癌癥發(fā)生機(jī)制的研究具有重要的科學(xué)意義和臨床價(jià)值。近年來,隨著熒光技術(shù)的快速發(fā)展,小分子熒光探針以其靈敏度高、操作簡便、體積小等特點(diǎn),已經(jīng)廣泛應(yīng)用于蛋白質(zhì)、核酸等重要生物分子的生物學(xué)和藥理學(xué)檢測中,對疾病機(jī)制探討、臨床診斷及藥物篩選等領(lǐng)域具有重要的意義。小分子熒光探針通常由兩部分組成：藥效基團(tuán)部分和熒光基團(tuán)部分。藥效基團(tuán)與目標(biāo)生物分子成高親和力結(jié)合,繼而熒光基團(tuán)通過激發(fā)而發(fā)射熒光以標(biāo)記蛋白質(zhì)。本文選取hERG鉀離子通道的抑制劑阿司咪唑和E-4031與通道主要結(jié)合部位的結(jié)構(gòu)作為藥效團(tuán),以香豆素類、萘二酰亞胺類、BD-C1、SBD-C1、Cy5類具有良好光學(xué)性質(zhì)的化合物作為熒光團(tuán),通過烷基鏈將兩者相連,設(shè)計(jì)合成了hERG通道的小分子熒光探針,并對其進(jìn)行了光學(xué)性質(zhì)的檢測、hERG鉀離子通道的親和力測試、細(xì)胞的熒光成像評價(jià)以及細(xì)胞毒性測試。結(jié)果顯示,所合成的hERG鉀通道的小分子熒光探針具有良好的光學(xué)性質(zhì)、較高的親和力,可選擇性的標(biāo)記hERG通道,并且其細(xì)胞毒性較小。綜上所述,通過本課題對hERG通道熒光探針的研究,以這些探針分子作為工具對hERG通道進(jìn)行標(biāo)記,將有可能為hERG鉀離子通道的生理病理研究提供新的幫助,若利用所得到的熒光探針進(jìn)一步進(jìn)行hERG通道抑制劑的篩選研究,將對新藥心臟毒性安全性評價(jià)具有重要的意義。此外,通過探針分子對腫瘤細(xì)胞的標(biāo)記也將為腫瘤的研究提供新的啟示。
[Abstract]:HERG ether-a-go-go related encoding fast activated delayed rectifier potassium current (IKR) potassium channel plays a key role in the repolarization of cardiac action potential. When hERG mutates or when hERG channels are blocked by drugs, it may cause hereditary or acquired long QT interval syndrome (LQTS). Long QT syndrome is caused by the blocking effect of different drugs on hERG channel. The safety of hER G channel has attracted much attention in drug development and clinical research. Therefore, hER G channel has been an important target for the screening of cardiac toxicity of drug candidates. In addition, preliminary studies showed that the expression of hERG was significantly up-regulated in some tumor cells, but not expressed or low in the corresponding normal cells of tumor cells. Furthermore, it was found that hERG channels were closely related to tumor differentiation, apoptosis and reproduction. This indicates that hERG may be a new biomarker for tumors. Therefore, the study of systemic physiology and pathology of hERG channel will be of great scientific significance and clinical value in evaluating the safety of cardiac toxicity and studying the mechanism of cancer in the preclinical study of new drugs. In recent years, with the rapid development of fluorescence technology, small molecular fluorescent probes have been widely used in the biological and pharmacological detection of proteins, nucleic acids and other important biomolecules because of their high sensitivity, simple operation and small size. It is of great significance to explore the mechanism of disease, clinical diagnosis and drug screening. Small molecular fluorescence probes usually consist of two parts: the effector group and the fluorescence group. The effector groups bind to target biomolecules with high affinity, and then fluorescence groups emit fluorescence to label proteins by excitation. In this paper, the structures of the main binding sites of hERG potassium channel inhibitor, Alimidazole and E-4031, were selected as pharmacophore groups. Coumarins and naphthalenediimide compounds with good optical properties were used as fluorescent groups. The small molecular fluorescence probes of hERG channel were designed and synthesized by linking them with alkyl chain. The optical properties of these probes were tested for their affinity, fluorescence imaging evaluation and cytotoxicity test. The results show that the synthesized hERG potassium channel fluorescent probe has good optical properties, high affinity, selective labeling of hERG channel, and its cytotoxicity is relatively small. In conclusion, the study of hERG channel fluorescent probes and the labeling of hERG channels with these probe molecules may provide new help for the physiological and pathological study of hERG potassium channels. If the fluorescent probe is used to screen hERG channel inhibitors further, it will be of great significance to evaluate the cardiac toxicity safety of new drugs. In addition, the labeling of tumor cells by probe molecules will provide new inspiration for tumor research.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R914

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