本文選題：pH響應(yīng) + 天然多糖鍵合藥��； 參考：《東北師范大學(xué)》2016年博士論文

【摘要】：隨著環(huán)境污染和食品安全等問題的日趨惡化,惡性腫瘤已成為威脅人類健康生命的疾病,其發(fā)病率和死亡率呈逐年上升的趨勢。目前,化療仍是腫瘤治療的最主要手段,但臨床經(jīng)常使用的小分子抗腫瘤藥物水溶性差,體內(nèi)循環(huán)周期短,代謝快,易產(chǎn)生多藥耐藥性,并且對(duì)腫瘤組織缺少靶向性。為了解決小分子藥物存在的諸多問題,通過物理包埋或化學(xué)鍵合的方式將小分子藥物與高分子結(jié)合而成的高分子納米藥物引起了廣泛的關(guān)注。本論文中,以具有良好生物相容性的天然多糖為載體骨架,采用化學(xué)鍵合的方式將小分子抗腫瘤藥物與載體材料結(jié)合,制備了一系列pH響應(yīng)的天然多糖鍵合藥,考察了藥物鍵合率對(duì)藥物傳輸?shù)挠绊?并進(jìn)一步研究了被動(dòng)和主動(dòng)靶向、及主動(dòng)靶向雙藥聯(lián)用對(duì)抗腫瘤效果的影響。主要研究內(nèi)容如下:(1)pH響應(yīng)的羥乙基淀粉-阿霉素鍵合藥(HES=DOX)體系選用羥乙基淀粉為基本骨架,利用氧化的羥乙基淀粉的醛基與阿霉素的氨基通過高效的席夫堿反應(yīng)合成酸敏感的HES=DOX,通過改變阿霉素投料摩爾比制備三種不同藥物鍵合率的阿霉素鍵合藥。所合成的三種天然多糖鍵合藥在水介質(zhì)中均可自組裝形成膠束,通過透射電鏡(TEM)和動(dòng)態(tài)光散射(DLS)對(duì)其形貌和粒徑進(jìn)行了考察。隨后對(duì)三種天然多糖鍵合藥的體外釋放、細(xì)胞毒性、細(xì)胞內(nèi)吞、體內(nèi)腫瘤抑制情況進(jìn)行研究。HES=DOX通過增強(qiáng)滲透與滯留效應(yīng)(EPR被動(dòng)靶向效應(yīng))可以到達(dá)腫瘤組織,并被細(xì)胞內(nèi)吞,在細(xì)胞內(nèi)低pH值的環(huán)境下快速裂解釋放出阿霉素,達(dá)到腫瘤抑制的目的。結(jié)果表明,三種天然多糖鍵合藥均可有效的降低小分子藥物的毒副作用,延長體內(nèi)循環(huán)時(shí)間,抑制腫瘤生長,且隨著藥物鍵合率的升高,腫瘤抑制能力增強(qiáng)。(2)主動(dòng)靶向的酸敏感的羥乙基淀粉-阿霉素鍵合藥(HES=DOX/cRGD)基于前面的研究,在天然多糖鍵合藥中引入主動(dòng)靶向分子,使天然多糖鍵合藥通過主動(dòng)靶向作用到達(dá)腫瘤組織,進(jìn)一步增加天然多糖鍵合藥在腫瘤組織的聚集,并通過與靶向受體結(jié)合進(jìn)入腫瘤細(xì)胞,在腫瘤細(xì)胞內(nèi)低pH情況下觸發(fā)釋放藥物。將帶有氨基的cRGD靶向分子和阿霉素分別通過席夫堿鍵連接到醛基化的羥乙基淀粉上。在2小時(shí)的細(xì)胞內(nèi)吞和胞內(nèi)藥物釋放實(shí)驗(yàn)中,αvβ3整合素過表達(dá)的A375細(xì)胞對(duì)HES=DOX/cRGD膠束具有更高的攝取。在體內(nèi)腫瘤抑制生長實(shí)驗(yàn)中,靶向天然多糖鍵合藥具有更高效的治療效果,并且通過免疫組化分析進(jìn)一步確定靶向天然多糖鍵合藥可以導(dǎo)致更多的腫瘤細(xì)胞凋亡,并且降低了對(duì)其他器官的毒副作用。(3)自靶向的酸敏感的葡聚糖-阿霉素鍵合藥(Dex-g-DOX)為了解決天然多糖鍵合藥膠束的藥物突釋和提高水溶性,以具有更好水溶性和易于功能化的天然多糖-葡聚糖為基礎(chǔ)骨架,制備酸敏感的葡聚糖基鍵合藥。葡聚糖和順式烏頭酸酐修飾的DOX通過簡單高效的縮合反應(yīng)制備酸敏感的天然多糖鍵合藥。值得注意的是,這兩個(gè)分子之間是通過酸裂解的酰胺鍵連接的。Dex經(jīng)體內(nèi)循環(huán)后選擇性聚集在肝臟部位,Dex-g-DOX實(shí)現(xiàn)在肝臟腫瘤部位的高強(qiáng)度聚集。由于Dex和DOX之間是通過酸響應(yīng)的酰胺鍵進(jìn)行連接的,所以Dex-g-DOX可以在酸性的內(nèi)涵體和溶酶體內(nèi)選擇釋放DOX,以增強(qiáng)DOX對(duì)腫瘤細(xì)胞的抑制能力。(4)具有主動(dòng)靶向的酸敏感的葡聚糖-阿霉素/硼替佐米協(xié)同鍵合藥(Dex-g-DOX/BTZ/cRGD)為了減少或消除單一藥物所存在的耐藥性,提高腫瘤抑制能力,將阿霉素(抑制核酸合成)和硼替佐米(抑制核因子-κB(NF-κB))分別通過酸敏感的席夫堿鍵和硼酯鍵與葡聚糖連接,并將能與αvβ3整合素受體相結(jié)合的靶向分子cRGD,也通過酸敏感的席夫堿鍵鍵合到葡聚糖上。所合成的靶向天然多糖鍵合藥膠束,通過靶向受體介導(dǎo)進(jìn)入腫瘤細(xì)胞,并在腫瘤細(xì)胞低pH的環(huán)境下釋放出抗腫瘤藥物,實(shí)現(xiàn)了藥物的協(xié)同治療。在本論文的研究中,得到了具有較好腫瘤治療效果的pH響應(yīng)的天然多糖鍵合藥。期望這種合成步驟簡單,治療效果顯著的天然多糖鍵合藥體系對(duì)高分子鍵合藥的臨床研究提供基礎(chǔ)實(shí)驗(yàn)依據(jù),為新型納米藥物的研發(fā)奠定基礎(chǔ)。
[Abstract]:With the worsening of environmental pollution and food safety, malignant tumor has become a disease that threatens human health. Its incidence and mortality are increasing year by year. At present, chemotherapy is still the most important means of cancer treatment, but the small molecular antitumor drugs often used in clinical use are poor in water solubility and short cycle cycle in the body. In order to solve the problems of small molecular drugs, a wide range of polymer nanoscale drugs are produced by combining physical encapsulation or chemical bonding with small molecular drugs and polymers. In this paper, a good biocompatibility is made in this paper. Natural polysaccharide was used as the carrier skeleton. A series of natural polysaccharide bonds with pH response were prepared by combining chemical bonding with the carrier materials. The effects of the drug bonding rate on the drug delivery were investigated, and the effects of passive and active targeting and the active target targeted double drugs were further studied. The main research contents are as follows: (1) hydroxyethyl starch in the hydroxyethyl starch and adriamycin bond (HES=DOX) system of pH responds to hydroxyethyl starch as the basic skeleton. The acid sensitive HES= DOX is synthesized by the reaction of the aldehyde group of oxidized hydroxyethyl starch and amido of adriamycin by high efficient Schiff base. Three different kinds of adriamycin feed ratio are prepared by changing the ratio of the dosage of adriamycin. The drug bond rate of adriamycin bond. The three kinds of natural polysaccharides can be assembled into a micelle in water medium. The morphology and particle size of the three compounds are investigated by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The release, cytotoxicity, endocytosis, and tumor inhibition of the three natural polysaccharide bonds are followed. .HES=DOX can reach tumor tissue by enhancing osmosis and retention effect (EPR passive targeting effect) and endocytosis by cells and release adriamycin in a low pH environment. The results show that the three natural polysaccharide bonds can effectively reduce the toxicity of small molecular drugs. Side effects, prolong the circulation time of the body, inhibit the growth of the tumor, and increase the tumor inhibition ability with the increase of the drug bonding rate. (2) the active target acid sensitive hydroxyethyl amyloid adriamycin bond (HES=DOX/cRGD) is based on the previous study, and the active target molecule is introduced into the natural polysaccharide bond, so that the natural polysaccharide bond is passed through the main drug. To reach the tumor tissue by moving target, further increase the accumulation of natural polysaccharide bond in tumor tissue, and enter into tumor cells by combining with the target receptor and trigger release drug under the low pH condition in the tumor cells. The cRGD target molecules with amino group and adriamycin are connected to the aldehyde based hydroxyethyl Lake respectively by the Schiff base bond. In 2 hours of endocytosis and intracellular drug release experiments, alpha v beta 3 integrin overexpressed A375 cells have higher uptake of HES=DOX/cRGD micelles. In vivo tumor inhibition growth experiments, targeted natural polysaccharide bonds have a more effective therapeutic effect, and the target nature is further determined by immunohistochemical analysis. Sugar bonds can cause more apoptosis of tumor cells and reduce the toxic and side effects to other organs. (3) the self targeting acid sensitive dextran doxorubicin bond (Dex-g-DOX) is used to solve the drug release and water solubility of natural polysaccharide bond micelles, with a better water-soluble and functional natural polysaccharide Portuguese. An acid sensitive glucan based bond is prepared based on chitosan. Dextran and CIS Aconitum modified DOX can be used to prepare acid sensitive natural polysaccharide bonds by a simple and efficient condensation reaction. It is worth noting that the two molecules are selectively aggregated in the liver through the acid cleavage of the amide linked.Dex through the body. In the dirty site, Dex-g-DOX is highly concentrated in the tumor site of the liver. Since Dex and DOX are linked by acid responsive amide bonds, Dex-g-DOX can release DOX in acid endosomes and enzymes to enhance the inhibitory ability of DOX to tumor cells. (4) acid sensitive dextran with active target. Mycophenin / bortezomib synergistic bond (Dex-g-DOX/BTZ/cRGD), in order to reduce or eliminate the drug resistance of a single drug, improves the tumor suppressive ability. Adriamycin (inhibition of nucleic acid synthesis) and bortezomib (inhibition of nuclear factor kappa B (NF- kappa B)) are connected by acid sensitive Schiff base and borate bonds to glucan respectively, and will be able to integrate with alpha v beta 3. The target molecule cRGD, combined with a peptide receptor, is also linked to dextran by acid sensitive Schiff base bond. The targeted natural polysaccharide binding micelles are introduced into tumor cells through targeted receptors, and antitumor drugs are released under the low pH environment of the tumor cells to achieve the synergistic treatment of drugs. In this paper, the study was carried out in this paper. The natural polysaccharide bond of pH response with better tumor treatment effect is obtained. It is expected that this synthetic procedure is simple. The natural polysaccharide bond system with significant therapeutic effect provides the basic experimental basis for the clinical study of polymer bond, and lays the foundation for the research and development of new nanoscale drugs.

【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R730.5;TQ464.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 徐立宏,張本山,高大維;羥乙基淀粉的制備與應(yīng)用[J];糧食與飼料工業(yè);2001年11期

，

本文編號(hào)：1835520