本文選題：光譜法 + 頭孢類藥物��； 參考：《河北大學(xué)》2017年碩士論文

【摘要】：近年來(lái),人類疾病的發(fā)生率逐年增加,各種藥物在生活中也起到了不容忽視的作用,為了更好的了解藥物的藥理、藥效和靶向運(yùn)輸,藥物與蛋白相互作用的研究課題已成為人們關(guān)注的重點(diǎn)。本文以胰蛋白酶和牛轉(zhuǎn)鐵蛋白為載體蛋白,用多種方法研究了硫酸頭孢匹羅、頭孢哌酮鈉、頭孢尼西鈉與蛋白的相互作用。研究?jī)?nèi)容分為五部分:第一章:綜述了幾種研究蛋白與小分子藥物相互作用的常用方法,并對(duì)胰蛋白酶和轉(zhuǎn)鐵蛋白進(jìn)行了簡(jiǎn)單的概述。對(duì)文章的立題思路、研究?jī)?nèi)容以及藥物與蛋白相互作用的展望進(jìn)行了簡(jiǎn)單的介紹。第二章:用多種光譜法和分子對(duì)接法研究了胰蛋白酶和硫酸頭孢匹羅之間的相互作用。結(jié)果表明,硫酸頭孢匹羅使胰蛋白酶發(fā)生熒光猝滅,猝滅機(jī)理為靜態(tài)猝滅,結(jié)合位點(diǎn)數(shù)接近于1。猝滅曲線表明色氨酸殘基和酪氨酸殘基都參加了反應(yīng)。同步熒光光譜法進(jìn)一步表明,二者主要的結(jié)合位點(diǎn)更接近于色氨酸殘基。對(duì)熱力學(xué)參數(shù)的計(jì)算表明,二者之間主要作用力為靜電引力,分子對(duì)接表明二者之間的作用力為靜電引力,其中還有氫鍵作用。胰蛋白酶與硫酸頭孢匹羅之間的距離小于7 nm,表明從胰蛋白酶到硫酸頭孢匹羅的能量轉(zhuǎn)移為非輻射能量轉(zhuǎn)移。熒光光譜法對(duì)Hill系數(shù)的研究表明,硫酸頭孢匹羅與胰蛋白酶之間存在負(fù)協(xié)同作用。第三章:利用熒光光譜法,同步熒光光譜法和紫外吸收光譜法研究了硫酸頭孢匹羅和牛轉(zhuǎn)鐵蛋白之間的相互作用。測(cè)定了二者之間的猝滅常數(shù)、結(jié)合常數(shù)、結(jié)合位點(diǎn)等。通過(guò)對(duì)熱力學(xué)參數(shù)的計(jì)算,確定了硫酸頭孢匹羅和牛轉(zhuǎn)鐵蛋白之間的主要作用力類型以靜電引力為主。通過(guò)對(duì)Hill系數(shù)的計(jì)算,得出了硫酸頭孢匹羅和牛轉(zhuǎn)鐵蛋白之間的協(xié)同作用為負(fù)協(xié)同作用。第四章:通過(guò)熒光猝滅法和同步熒光光譜法對(duì)頭孢哌酮鈉與牛轉(zhuǎn)鐵蛋白的相互作用機(jī)理進(jìn)行了研究。結(jié)果表明,兩種方法用相同的公式計(jì)算得到的二者的猝滅機(jī)理、結(jié)合位點(diǎn),相互作用力以及協(xié)同作用基本一致。并且通過(guò)對(duì)Δλ=60 nm和λex=295 nm的數(shù)據(jù)進(jìn)行比較,得出同步熒光光譜法較之于傳統(tǒng)熒光光譜法有更高的靈敏度和準(zhǔn)確度。因此,同步熒光光譜法極大地豐富了小分子配體與蛋白之間相互作用機(jī)理的研究方法。第五章:實(shí)驗(yàn)在pH=7.4緩沖溶液條件下,研究了不同溫度下頭孢尼西鈉與牛轉(zhuǎn)鐵蛋白的相互作用。結(jié)果表明,牛轉(zhuǎn)鐵蛋白-頭孢尼西鈉體系中,主要的作用方式為生成了新的復(fù)合物的靜態(tài)猝滅,二者的結(jié)合位點(diǎn)數(shù)為1。由熱力學(xué)參數(shù)可知頭孢尼西鈉與牛轉(zhuǎn)鐵蛋白主要通過(guò)靜電引力結(jié)合。紫外吸收光譜法和同步熒光光譜法都證明了頭孢尼西鈉使得牛轉(zhuǎn)鐵蛋白氨基酸極性增大,疏水性減小。
[Abstract]:In recent years, the incidence of human diseases has increased year by year, and various drugs have played an important role in life. In order to better understand the pharmacology, efficacy and targeted transportation of drugs, The study of drug-protein interaction has become the focus of attention. Using trypsin and bovine transferrin as carrier proteins, the interaction of cefpirosine sulfate, cefoperazone sodium and cefnesian sodium with protein was studied by various methods. The research contents are divided into five parts: chapter 1: the common methods of studying the interaction between protein and small molecule drugs are reviewed, and the trypsin and transferrin are briefly summarized. The idea, research content and prospect of drug-protein interaction are briefly introduced in this paper. In chapter 2, the interaction between trypsin and cefpirome sulfate was studied by multiple spectral and molecular docking methods. The results showed that cefpirosine sulfate caused fluorescence quenching of trypsin, the quenching mechanism was static quenching, and the binding site number was close to 1. The quenching curves showed that both tryptophan residues and tyrosine residues participated in the reaction. Synchronous fluorescence spectroscopy further showed that the binding sites of the two were closer to tryptophan residues. The calculation of thermodynamic parameters shows that the main force between them is electrostatic force, and the molecular docking shows that the force between them is electrostatic force, among which there is hydrogen bond. The distance between trypsin and cefpirosine sulfate is less than 7 nm, which indicates that the energy transfer from trypsin to cefpirome sulfate is non-radiation energy transfer. The study of Hill coefficient by fluorescence spectroscopy showed that there was a negative synergistic effect between cefopirol sulfate and trypsin. In chapter 3, the interaction between cefpirosine sulfate and bovine transferrin was studied by fluorescence spectroscopy, synchronous fluorescence spectroscopy and ultraviolet absorption spectrometry. The quenching constants and binding sites were determined. Based on the calculation of thermodynamic parameters, the main force between cefopiro sulfate and bovine transferrin was determined to be electrostatic force. By calculating the Hill coefficient, the synergism between cefopirol sulfate and bovine transferrin was found to be negative. Chapter 4: the interaction mechanism between cefoperazone sodium and bovine transferrin was studied by fluorescence quenching and synchronous fluorescence spectrometry. The results show that the quenching mechanism, binding site, interaction force and synergism between the two methods calculated by the same formula are basically the same. By comparing the data of 螖 位 ~ (60) nm and 位 ex=295 nm, it is concluded that the synchronous fluorescence spectrum method has higher sensitivity and accuracy than the traditional fluorescence spectrum method. Therefore, synchronous fluorescence spectroscopy has greatly enriched the mechanism of interaction between small molecular ligands and proteins. Chapter 5: the interaction between cefnesian sodium and bovine transferrin at different temperatures in pH=7.4 buffer solution was studied. The results showed that the main action mode of bovine transferin-cefnesian sodium system was the static quenching of the formation of new complexes, the binding site of which was 1. Thermodynamic parameters show that cefnesian sodium binds to bovine transferrin mainly by electrostatic force. Both UV absorption and synchronous fluorescence spectra showed that cefnesian sodium increased amino acid polarity and decreased hydrophobicity of bovine transferrin.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R96;O657.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉景陶;劉映雪;;計(jì)算機(jī)輔助藥物設(shè)計(jì)在分子對(duì)接中的應(yīng)用[J];科技創(chuàng)新與應(yīng)用;2016年34期

2 陳晉瑩;桑梓苔;廖子龍;;運(yùn)用計(jì)算機(jī)分子對(duì)接技術(shù)初步探究糧食中玉米赤霉烯酮及其降解產(chǎn)物的雌激素效應(yīng)[J];糧食儲(chǔ)藏;2016年02期

3 Vishwas D.Suryawanshi;Laxman S.Walekar;Anil H.Gore;Prashant V.Anbhule;Govind B.Kolekar;;Spectroscopic analysis on the binding interaction of biologically active pyrimidine derivative with bovine serum albumin[J];Journal of Pharmaceutical Analysis;2016年01期

4 郭明;范文翔;吳榮暉;;不同血清蛋白輸運(yùn)厚樸酚的差異性及模型適用度分析[J];中國(guó)藥學(xué)雜志;2013年24期

5 郭明;詹敏忠;魯小旺;范文翔;;金銀花活性成分綠原酸與不同蛋白質(zhì)相互作用機(jī)制的比較分析研究[J];中國(guó)中藥雜志;2013年16期

6 郭明;魯小旺;冉曉云;胡潤(rùn)淮;;轉(zhuǎn)鐵蛋白轉(zhuǎn)運(yùn)培氟沙星的分子作用機(jī)制研究[J];藥學(xué)學(xué)報(bào);2012年11期

7 劉保生;楊超;王晶;薛春麗;呂運(yùn)開(kāi);;硫酸頭孢匹羅與牛血清白蛋白結(jié)合反應(yīng)的發(fā)光機(jī)理[J];發(fā)光學(xué)報(bào);2011年03期

8 梁彥秋;臧樹(shù)良;趙雪;;頭孢他啶與牛血清白蛋白(BSA)的相互作用[J];光譜實(shí)驗(yàn)室;2009年06期

9 姜虹;安普麗;蔣曄;;藥物與蛋白質(zhì)相互作用研究方法的進(jìn)展[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2007年06期

10 沈星燦,梁宏,何錫文,王新省;圓二色光譜分析蛋白質(zhì)構(gòu)象的方法及研究進(jìn)展[J];分析化學(xué);2004年03期

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