本文選題：苯丙氨酸　切入點：血管緊張素轉(zhuǎn)化酶抑制肽　出處：《廣東藥科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：血管緊張素轉(zhuǎn)化酶(ACE)在血壓調(diào)節(jié)中的關(guān)鍵性作用,使其成為治療高血壓疾病的理想靶點。ACE抑制肽憑借其安全、易吸收等優(yōu)點受到了廣泛關(guān)注。本論文采用以苯丙氨酸為碳端的ACE抑制二、三肽進(jìn)行三維定量構(gòu)效關(guān)系(3D-QSAR)研究并建立相關(guān)模型,根據(jù)所建模型篩選并合成具有良好ACE抑制活性的三肽并進(jìn)行體外活性驗證,同時采用分子動力學(xué)模擬和二維核磁共振實驗相結(jié)合的方法,研究新型ACE抑制肽的分子作用機(jī)制,并給出微觀解釋。本文的主要內(nèi)容由以下四部分組成:第一部分,收集已報道的53種ACE抑制二、三肽進(jìn)行3D-QSAR CoMFA/CoMSIA以及分子對接研究,建立了具有良好預(yù)測能力的CoMFA/CoMSIA模型,并獲得抑制肽分子與ACE受體之間可能的結(jié)合模式。同時根據(jù)已建立的模型篩選并合成了四種三肽Gly-Glu-Phe(GEF),Val-Glu-Phe(VEF),Val-Arg-Phe(VRF)以及Val-Lys-Phe(VKF)進(jìn)行體外活性實驗,對模型進(jìn)行進(jìn)一步的驗證。第二部分,采用改良的紫外分光光度法對第一部分中篩選并合成的四種三肽GEF,VEF,VRF以及VKF進(jìn)行體外活性實驗驗證。其中,GEF的CoMFA/CoMSIA模型預(yù)測值均與實驗值較為接近,誤差分別為4.49%和7.14%;VEF的Co MSIA模型預(yù)測值與實驗值的相對誤差最小,僅為1.08%;VRF的CoMFA模型預(yù)測值與實驗值之間的相對誤差最大,為9.87%,但是兩者仍然屬于同一個數(shù)量級。上述四種三肽的CoMFA/CoMSIA模型預(yù)測值均接近與實驗活性值,實驗結(jié)果進(jìn)一步證明了所建模型具有良好的預(yù)測能力及準(zhǔn)確性。第三部分,采用分子動力學(xué)模擬與2D 1H-1H-NOESY圖譜相結(jié)合的方法,研究生物活性水溶液GEF-H2O體系和小肽變性條件下的GEF-DMSO體系中GEF分子的構(gòu)象變化以及分子間的弱相互作用。結(jié)果表明GEF分子在水溶液中主要以伸展構(gòu)象存在,折疊構(gòu)象和伸展構(gòu)象之間可以快速相互轉(zhuǎn)換,而在DMSO溶液中主要以折疊構(gòu)象存在,較少存在伸展構(gòu)象。2D 1H-1H-NOESY圖譜的得到的結(jié)果與分子動力學(xué)模擬結(jié)論一致,理論和實驗的結(jié)果得到很好的吻合。第四部分,本章采用分子動力學(xué)研究了GEF分子分別與ACE的C-結(jié)構(gòu)域(C-ACE)、N-結(jié)構(gòu)域(N-ACE)之間相互作用的機(jī)制。研究發(fā)現(xiàn)GEF分子在C-ACE活性區(qū)域的構(gòu)象較在N-ACE活性區(qū)域的構(gòu)象更舒展,C-ACE的Lys475氨基酸殘基對配體與受體之間的結(jié)合影響最大,而N-ACE中影響最大的是Lys489氨基酸殘基。通過配位作用分析發(fā)現(xiàn),GEF分子與Zn離子之間的相互作用是維持配體-受體復(fù)合物穩(wěn)定的主要因素。本論文運用理論與實驗相結(jié)合的方法建立了一套篩選高活性ACE抑制肽及揭示分子作用機(jī)制的研究體系。這一體系包括:建立ACE抑制肽的數(shù)學(xué)模型;篩選并合成具有較高抑制活性的肽;體外活性實驗驗證模型的準(zhǔn)確性;分子動力學(xué)模擬從微觀角度解釋活性分子結(jié)構(gòu)與活性之間的關(guān)系。此方法對研究和開發(fā)新型ACE抑制肽具有一定的理論指導(dǎo)意義。
[Abstract]:The key role of angiotensin converting enzyme (ACE) in the regulation of blood pressure makes it an ideal target for the treatment of hypertension. The advantages of easy absorption have attracted much attention. In this paper, the three-dimensional quantitative structure-activity relationship (3D-QSAR) was studied by using phenylalanine (phenylalanine) as carbon terminal ACE to inhibit dipeptide and tripeptide, and the relevant models were established. According to the established model, tripeptides with good ACE inhibitory activity were selected, synthesized and verified in vitro. At the same time, the molecular mechanism of the novel ACE inhibitory peptides was studied by means of molecular dynamics simulation and two-dimensional nuclear magnetic resonance (NMR). The main contents of this paper are as follows: in the first part, we collect 53 kinds of ACE inhibitory dipeptide and tripeptide to study 3D-QSAR CoMFA/CoMSIA and molecular docking, and establish a CoMFA/CoMSIA model with good prediction ability. The possible binding patterns between inhibitory peptide molecules and ACE receptors were obtained. Four tripeptide Gly-Glu-Phegnon Val-Glu-Phegnon VRFs and Val-Lys-Phe-Phe-VKFs were screened and synthesized according to the established model. Modified UV spectrophotometry was used to verify the in vitro activity of four tripeptide GEFFV VRF and VKF, which were selected and synthesized in the first part. The predicted values of CoMFA/CoMSIA model were all close to the experimental values. The relative error between the predicted value and experimental value of Co MSIA model with errors of 4.49% and 7.14 respectively is the smallest, and the relative error between the predicted value of CoMFA model and the experimental value of 1.08VRF is the largest. The predicted values of the CoMFA/CoMSIA model of the above four tripeptides are all close to the experimental activity values. The experimental results further prove that the model has good predictive ability and accuracy. The method of combining molecular dynamics simulation with 2D 1H-1H-NOESY map was used. The conformational changes and weak intermolecular interactions of GEF molecules in bioactive aqueous solution GEF-H2O system and GEF-DMSO system with small peptide denaturation were studied. The results show that GEF molecules mainly exist in extensional conformation in aqueous solution. The folding conformation and extensional conformation can be converted rapidly, while in DMSO solution, the folding conformation mainly exists in the form of folded conformation, and the results obtained from the extensional conformation .2D 1H-1H-NOESY are in agreement with the results obtained by molecular dynamics simulation. The theoretical and experimental results are in good agreement. Part 4th, In this chapter, molecular dynamics was used to study the mechanism of interaction between GEF molecule and C-ACE-N-ACE-domain of ACE. It was found that the conformation of GEF molecule in C-ACE active region was more extensive than that in N-ACE active region. The amino acid residues of Lys475 have the greatest influence on the binding between ligand and receptor. In N-ACE, the amino acid residues of Lys489 were the most influential. The interaction between Lys489 molecules and Zn ions was found to be the main factor to maintain the stability of ligand receptor complexes. In this paper, the theoretical and experimental phase was used. A set of research systems for screening highly active ACE inhibitory peptides and revealing the molecular mechanism of action were established by the combined method. The system includes: establishing a mathematical model of ACE inhibitory peptides; The peptides with high inhibitory activity were screened and synthesized, and the accuracy of the model was verified by in vitro activity experiment. Molecular dynamics simulation explains the relationship between active molecular structure and activity from the microcosmic point of view. This method has a certain theoretical significance for the research and development of novel ACE inhibitory peptides.
【學(xué)位授予單位】：廣東藥科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R945

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉龍珠;賀熙;徐曼;于曉江;臧偉進(jìn);;高血壓免疫機(jī)制的研究進(jìn)展[J];生理科學(xué)進(jìn)展;2016年04期

2 周海玲;馬麟;易智彪;;基于三種體外抗氧化方法對白木香種子抗氧化能力的研究[J];中國醫(yī)藥導(dǎo)報;2016年22期

3 王月;韓梅玲;成麗娟;孫尚文;刁玉濤;;用統(tǒng)計軟件計算腎小球系膜細(xì)胞對順鉑的半數(shù)抑制濃度[J];預(yù)防醫(yī)學(xué)論壇;2016年06期

4 吳憲明;孫躍民;;焦慮抑郁與高血壓[J];中華高血壓雜志;2016年02期

5 孫英賢;;高血壓治療新藥研究進(jìn)展[J];中國實用內(nèi)科雜志;2015年04期

6 吳堅;薛曉燕;王麗芳;郭小華;肖林林;;分子對接方法應(yīng)用與發(fā)展[J];亞太傳統(tǒng)醫(yī)藥;2013年12期

7 熊浩明;魏柏青;魏榮杰;楊寧海;祁美英;靳娟;金泳;何建;代瑞霞;;用SPSS軟件計算鼠疫菌半數(shù)致死量(LD_(50))[J];中國人獸共患病學(xué)報;2013年11期

8 彭劍秋;劉靜;管驍;;一種新的氨基酸描述符SVHEHS在生物活性肽QSAR中的應(yīng)用研究[J];食品科學(xué);2012年07期

9 劉靜;彭劍秋;管驍;;基于多元線性回歸的血管緊張素轉(zhuǎn)化酶抑制肽定量構(gòu)效關(guān)系建模研究[J];分析科學(xué)學(xué)報;2012年01期

10 仝艷;李曉曉;李曉飛;;介紹一款優(yōu)秀的分子圖形軟件Chimera[J];化工時刊;2011年10期

，

本文編號：1621679