本文選題：熒光探針　切入點(diǎn)：Cu~(2+)_L-組氨酸　出處：《中國(guó)科學(xué)技術(shù)大學(xué)》2014年博士論文

【摘要】：熒光分子探針分析技術(shù)是一種重要的分子檢測(cè)技術(shù),具有靈敏度高、選擇性好、檢測(cè)方便、檢測(cè)限低等特點(diǎn)。近幾十年來,熒光分子探針技術(shù)在環(huán)境科學(xué)、醫(yī)藥以及生命科學(xué)等領(lǐng)域被廣為應(yīng)用。 在論文中我們發(fā)展了一種新型的熒光可控探針Ac-SAACQ-Gly-Gly-Gly-Lys (FITC),該探針能被用于連續(xù)的選擇性在體外和活細(xì)胞中檢測(cè)Cu2+和L-組氨酸。分子探針結(jié)構(gòu)中包含的Gly-Gly-Gly-Lys能夠增加探針的水溶性和細(xì)胞滲透性；分子探針結(jié)構(gòu)中SAACQ部分能夠與Cu2+發(fā)生螯合作用；分子探針的熒光信號(hào)則來自于異硫氰酸熒光素(FITC),當(dāng)探針分子的SAACQ部分與Cu2+結(jié)合以后由于PET現(xiàn)象的出現(xiàn)導(dǎo)致熒光素的熒光發(fā)生淬滅。最終我們成功實(shí)現(xiàn)了體外和活細(xì)胞內(nèi)Cu2+以及L-組氨酸的連續(xù)性檢測(cè)。 定點(diǎn)標(biāo)記蛋白在檢測(cè)蛋白的表達(dá)、組合、轉(zhuǎn)錄,理解細(xì)胞內(nèi)部隨時(shí)間空間的變化具有重要的意義。在過去的十年中熒光蛋白已經(jīng)廣泛的應(yīng)用到標(biāo)記相應(yīng)的fusion蛋白,并且具有很好的分辨率。但是它們具有高分子量以及對(duì)細(xì)胞環(huán)境遲鈍的缺點(diǎn)。而化學(xué)探針則具有高標(biāo)記的特點(diǎn)。 在論文中我們通過合理改進(jìn)傳統(tǒng)的基于1,2-氨基硫醇和2-氰基苯并噻唑(CBT)的氰基的縮合反應(yīng)中的底物,將該反應(yīng)應(yīng)用到分子成像。我們的研究體系結(jié)合了巰基與馬來酰亞胺之間的親核加成反應(yīng)和CBT的-CN與半胱氨酸(Cys)的N端之間的縮合反應(yīng),最后成功的對(duì)雞蛋膜上蛋白質(zhì)的Cys殘基的進(jìn)行熒光標(biāo)記。 在臨床上癌癥多藥耐藥(MDR)的存在是癌癥病人實(shí)施化學(xué)治療失敗的主要原因。多藥耐藥不僅對(duì)接觸腫瘤細(xì)胞的藥物產(chǎn)生耐藥,而且對(duì)一些未曾接觸、與之化學(xué)結(jié)構(gòu)和作用機(jī)制完全不同的藥物也產(chǎn)生交叉耐藥,因此這給癌癥的治療帶來了很大的障礙。而納米載藥體系的出現(xiàn)為克服MDR帶來重大的突破。 在論文中我們合理的設(shè)計(jì)了四種抗癌藥物1,2,3以及對(duì)照化合物3-Scr。其中,2是化療藥物阿霉素(DOX)的衍生物。其他化合物則是紫杉醇(taxol)的衍生物。1,2,3都包含一段Arg-Val-Arg-Arg (RVRR)的肽鏈,該肽鏈能夠被furin酶特異性的識(shí)別和剪切。另外,RVRR肽鏈片段使底物具有很好的水溶性以及細(xì)胞滲透性。當(dāng)藥物分子被furin酶剪切或者還原劑還原以后,縮合反應(yīng)就會(huì)發(fā)生形成多聚體,進(jìn)而發(fā)生自組裝形成納米顆粒,同時(shí)增加細(xì)胞內(nèi)化療藥物的有效濃度。
[Abstract]:Fluorescence molecular probe analysis is an important molecular detection technology with high sensitivity, good selectivity, easy detection and low detection limit.In recent decades, fluorescent molecular probe technology has been widely used in environmental science, medicine and life sciences.In this paper, we have developed a novel fluorescent controllable probe, Ac-SAACQ-Gly-Gly-Gly-Lys FITCX, which can be used for the continuous selective detection of Cu2 and L-histidine in vitro and in living cells.The Gly-Gly-Gly-Lys contained in the molecular probe structure can increase the water solubility and cell permeability of the probe, and the SAACQ part of the molecular probe structure can chelate with Cu2.The fluorescence signal of the probe comes from the fluorescence of fluorescein isothiocyanate (FITC). When the SAACQ part of the probe molecule binds to Cu2, the fluorescence of fluorescein is quenched due to the PET phenomenon.Finally, we successfully achieved in vitro and in vivo Cu2 and L-histidine continuity detection.It is of great significance to detect the expression, combination, transcription and understanding of the changes in cell interior with time and space.In the past decade, fluorescent proteins have been widely used to label the corresponding fusion proteins and have good resolution.But they have the disadvantages of high molecular weight and being slow to the cell environment.Chemical probes are characterized by high labeling.In this paper, we apply this reaction to molecular imaging by reasonably improving the substrate in the traditional condensation reaction of cyanide based on 1- (2-aminothiol) and 2-cyanobenzothiazolium (CBT).Our system combines the nucleophilic addition reaction between mercapto and maleimide and the condensation reaction between the N-terminal of CBT and cysteine cysteine. Finally, the fluorescent labeling of the Cys residue of protein on egg membrane is carried out successfully.The presence of MDR is the main reason for the failure of chemotherapy in cancer patients.Multidrug resistance (MDR) is not only resistant to drugs in contact with tumor cells, but also cross-resistant to drugs that have not been exposed to and have completely different chemical structure and mechanism of action. Therefore, this has brought great obstacles to the treatment of cancer.The emergence of nano-drug-loaded system has brought about a great breakthrough in overcoming MDR.In this paper, we reasonably designed four anticancer drugs 1H 2C 3 and the control compound 3 Scr3.It is a derivative of doxorubicin (DOX), a chemotherapeutic drug.The other compounds are taxol taxol. the derivative of taxol.1ON2O3 contains a peptide chain of Arg-Val-Arg-Arg rvr. the peptide chain can be specifically identified and cut by furin enzyme.In addition, the RVRR peptide chain fragment makes the substrate have good water solubility and cell permeability.When the drug molecule is shredded by furin enzyme or reduced by reducing agent, the condensation reaction will take place to form a polymer, and then self-assemble to form nanoparticles, and increase the effective concentration of chemotherapeutic drugs in cells at the same time.
【學(xué)位授予單位】：中國(guó)科學(xué)技術(shù)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R914;O657.3

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 黃建艷;湯順清;蔡繼業(yè);;納米靶向技術(shù)在智能釋藥系統(tǒng)的應(yīng)用[J];材料導(dǎo)報(bào);2007年S3期

2 盤杰;陳軍;王治平;黃宇華;趙建江;;兩種粒徑量子點(diǎn)在人舌鱗癌細(xì)胞中雙色熒光成像的應(yīng)用研究[J];廣東牙病防治;2012年05期

3 劉t犇，

本文編號(hào)：1726962