【摘要】：隨著納米科學(xué)技術(shù)的發(fā)展,納米藥物遞送體系的應(yīng)用為腫瘤的診斷和治療提供了有效解決途徑。納米載藥體系可將藥物和成像試劑集成于納米顆粒中,通過(guò)增強(qiáng)型滲透效應(yīng)或主動(dòng)靶向作用有針對(duì)性地遞送到腫瘤組織,提高治療效果和減少對(duì)正常組織的毒副作用。在眾多納米載藥體系中,由于生物大分子納米載藥體系具備良好的生物相容性與生物可降解性,因此可避免其在體內(nèi)長(zhǎng)期累積所導(dǎo)致的毒性。基于生物可降解性納米材料的優(yōu)點(diǎn),本論文圍繞改善腫瘤診療效果這一目標(biāo),開發(fā)了基于牛血清白蛋白、藻藍(lán)蛋白和DNA的生物大分子納米載藥體系,將其應(yīng)用于腫瘤的靶向治療、影像示蹤、扭轉(zhuǎn)耐藥和放療/化療協(xié)同治療等。主要的研究工作如下:第一章介紹了納米藥物在腫瘤診療中的發(fā)展現(xiàn)狀,概述了生物大分子納米載體的類別、優(yōu)勢(shì)及其在腫瘤診斷和治療中的應(yīng)用。在第二章中,為了提高放療效果,合成了葉酸靶向的白蛋白納米載藥體系,并負(fù)載有機(jī)硒衍生物PSeD,構(gòu)建了兼具腫瘤靶向性與生物相容性的白蛋白納米載藥體系(FA-BSANP@Se),其能聯(lián)合X射線能大幅度提高宮頸癌Hela細(xì)胞內(nèi)的活性氧水平,使其發(fā)生大規(guī)模DNA損傷,從而級(jí)聯(lián)激活G2/M周期阻滯和凋亡信號(hào),最終抑制Hela細(xì)胞克隆群落的形成。為了克服異源性BSANPs可能對(duì)人體造成的免疫源性,在第三章中制備了無(wú)免疫源性的新型蛋白納米載藥體系—藻藍(lán)蛋白納米粒子(Phycocyanin nanoparticles,PCNPs)作為阿霉素(doxorubicine,DOX)的新型納米載體,并裝載靶向分子葉酸(folate,FA),探究其在扭轉(zhuǎn)腫瘤細(xì)胞對(duì)阿霉素耐藥的作用及相關(guān)機(jī)制。藥代動(dòng)力學(xué)研究表明,PCNPs明顯延長(zhǎng)了DOX的血液半衰期,在酸性溶酶體環(huán)境中釋放DOX。FA-PCNP@DOX通過(guò)FA/FARs介導(dǎo)的靶向作用識(shí)別腫瘤耐藥細(xì)胞RHep G2,通過(guò)上調(diào)胞內(nèi)活性氧水平引起線粒體斷裂,繼而阻斷ATP依賴性ABC轉(zhuǎn)運(yùn)蛋白的活性,顯著提高了DOX在RHepG2細(xì)胞內(nèi)的吸收和停留效率,最終引發(fā)RHepG2細(xì)胞凋亡�；铙w成像實(shí)驗(yàn)證明,FA-PCNP@DOX能靶向腫瘤區(qū)域,在有效抑制瘤體增殖的同時(shí)降低DOX引起的肝、肺、腎毒性。進(jìn)一步地,為了改善金屬配合物的血液相容性和靶向抗腫瘤效果,在第四章中構(gòu)建了cRGD靶向藻藍(lán)蛋白納米粒子作為釕多吡啶配合物(Ruthenium polypyridyl complexes,RuPOP)的載體,發(fā)現(xiàn)cRGD-PCNP@Ru能提高RuPOP的親水性,并阻止RuPOP和血紅細(xì)胞相互作用,從而改善了RuPOP的血液相容性。通過(guò)對(duì)一系列宮頸癌細(xì)胞Caski、Siha和Hela細(xì)胞進(jìn)行活性篩選,發(fā)現(xiàn)cRGD-PCNP@Ru能通過(guò)RGD-整合素介導(dǎo)的靶向作用識(shí)別整合素高表達(dá)的宮頸癌細(xì)胞Caski,提高了RuPOP在Caski腫瘤球內(nèi)部的累積,從而提高了RuPOP對(duì)Caski腫瘤球增殖的抑制作用。為了提高納米載藥體系和對(duì)金屬配合物的負(fù)載效率及體內(nèi)抗腫瘤活性,在第五章中構(gòu)建了Biotin靶向DNA cages納米載藥體系作為金屬配合物RuPOP的載體(Bio-Cage@Ru),研究其在提高RuPOP抗腫瘤活性和降低毒性方面的潛能。Bio-cage作為具有腫瘤靶向性的四面體籠型結(jié)構(gòu),通過(guò)生物素/受體介導(dǎo)的靶向作用增強(qiáng)了RuPOP對(duì)不同腫瘤細(xì)胞和正常細(xì)胞選擇性,實(shí)現(xiàn)了良好的抗腫瘤效果。Bio-cage@Ru加速RuPOP定位至細(xì)胞核,并在DNase I的作用下裂解DNA結(jié)構(gòu),促進(jìn)RuPOP的釋放。在荷瘤裸鼠模型中,Bio-cage@Ru提高了RuPOP在腫瘤區(qū)域的累積,從而明顯提高了其體內(nèi)抗腫瘤效果。此外,Bio-cage@Ru明顯降低了RuPOP所引起的心、肝、肺、腎毒性,整體實(shí)現(xiàn)了高效、低毒的目標(biāo)。在第五章中對(duì)全文工作進(jìn)行了總結(jié),并對(duì)未來(lái)的研究方向提出了展望。本論文工作的開展為癌癥治療中出現(xiàn)的放療、化療效果差、毒副作用明顯以及腫瘤耐藥等問(wèn)題提供了解決方法。
[Abstract]:With the development of nano science and technology, the application of nano drug delivery system provides an effective way for diagnosis and treatment of tumor. The nano drug loading system can integrate the drug and the imaging agent into the nano particles, and is targeted to the tumor tissue through the enhanced osmotic effect or the active targeting effect, thereby improving the treatment effect and reducing the toxic and side effects on the normal tissues. In many nano-drug delivery systems, since the biological macromolecule nano-drug carrier system has good biocompatibility and biodegradability, toxicity caused by long-term accumulation in vivo can be avoided. Based on the advantages of biodegradable nano materials, this paper aims at improving the diagnosis and treatment effect of tumor, and develops a biological macromolecule nano medicine loading system based on bovine serum albumin, phycoerythrin and DNA, which is applied to targeted therapy and image tracing of tumors. torsion resistance and radiotherapy/ chemotherapy co-treatment, etc. The main research work is as follows: The first chapter introduces the development status of nano-drugs in tumor diagnosis and treatment, and summarizes the categories, advantages and applications of biological macromolecule nano-carriers in tumor diagnosis and treatment. In the second chapter, in order to improve the effect of radiotherapy, folic acid-targeted albumin nano-drug delivery system was synthesized and the organic selenium derivative PSeD was loaded, and the albumin nano-drug delivery system (FA-BSANP@Se) with tumor targeting and biocompatibility was constructed. The combined X-ray can greatly improve the active oxygen level in the uterine cervix cancer cell and cause large-scale DNA damage, so that the G2/ M cycle block and the apoptosis signal are cascade activated, and finally the formation of the cell clone community of the cervical cancer cell is inhibited. In order to overcome the possible immune source of heterogenic BSANPs to human body, a new type of nano-carrier of phycoerythrin nanoparticles (PCNPs) as adriamycin (DOX) was prepared in the third chapter, and targeted molecular folic acid (Folate) was loaded. FA) to explore its role in reversing adriamycin resistance in tumor cells and its related mechanisms. Pharmacokinetic studies indicate that PCNPs significantly prolong the blood half-life of DOX, release DOX.FA-PCNP@DOX in acidic lysosomal environments to recognize tumor-resistant cells RHep G2 through FA/ FARs-mediated targeting, and cause mitochondrial breakage by up-regulation of intracellular reactive oxygen levels, and then blocking the activity of the ATP-dependent ABC transporter, remarkably improving the absorption and staying efficiency of DOX in the RHepG2 cells, and finally inducing the apoptosis of the RHepG2 cells. In vivo imaging experiments prove that the FA-PCNP@DOX can target tumor regions and reduce the toxicity of liver, lung and kidney caused by DOX while effectively inhibiting the proliferation of tumor bodies. Further, in order to improve the blood compatibility and targeted anti-tumor effect of metal complexes, we constructed the vector of cGVHD-targeting phycoerythrin nanoparticles as the carrier of polychrome complex (RuPOP) in the fourth chapter, and found that the cRGD-PCNP@Ru can improve the hydrophilicity of RuPOP. and prevents the interaction of RuPOP and red blood cells, thereby improving the blood compatibility of RuPOP. By carrying out active screening on a series of cervical cancer cells Caski, Siha, and Caski cells, it is found that the cRGD-PCNP@Ru can recognize the high-expression cervical cancer cell Caski through the targeting action mediated by the caspase-whole, and the accumulation of the RuPOP in the Caski tumor ball is improved, thereby improving the inhibiting effect of RuPOP on the proliferation of Caski tumor balls. In order to improve the loading efficiency and anti-tumor activity of the nano-drug delivery system and the metal complex, Biotin-targeted DNA images nano-drug delivery system was constructed as the carrier (Bio-Cage@Ru) of the metal complex RuPOP in chapter V to study its potential in improving the antitumor activity of RuPOP and reducing toxicity. Bio-Cage, as a tetrahedral cage type structure with tumor targeting property, enhances the selectivity of RuPOP to different tumor cells and normal cells by biotin/ receptor-mediated targeting action, realizes good anti-tumor effect .Bio-cage@Ru, accelerates the localization of RuPOP to the nucleus, and the DNA structure is cleaved under the action of DNase I to promote the release of RuPOP. In the nude mouse model of tumor-bearing mice, the Bio-cage@Ru increased the accumulation of RuPOP in the tumor area, thus improving the anti-tumor effect in vivo. In addition, the Bio-cage@Ru obviously reduces the toxicity of heart, liver, lung and kidney caused by RuPOP, and achieves the goal of high efficiency and low toxicity. In the fifth chapter, the thesis summarizes the work of the whole thesis, and puts forward the future research direction. The work of this thesis provides a solution to the problems of radiation therapy, poor chemotherapy effect, obvious toxic and side effect and drug resistance in cancer treatment.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R91;R730
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本文編號(hào)：2252696