【摘要】：腫瘤納米靶向化療(nanotarget chemotherapy)是指將化療藥物利用納米技術(shù)靶向輸送到腫瘤部位,以在發(fā)揮最大的療效同時(shí)減少化療藥物的毒副作用,而腫瘤納米靶向治療的關(guān)鍵在于納米載體材料。良好的納米載體能夠保護(hù)藥物在體內(nèi)循環(huán)不被過早的降解,而在到達(dá)靶部位后可順利釋藥,同時(shí)自身降解代謝為無毒代謝物。因此開發(fā)新型良好的納米藥物載體材料存在一定的挑戰(zhàn),且具有非常重要的意義。本課題基于腫瘤細(xì)胞的微環(huán)境及相關(guān)研究,合成了一種新型的可生物降解的高分子載體材料---交聯(lián)硫辛酸。此材料成分單純,制備工藝簡(jiǎn)單可控,穩(wěn)定性較好,毒性較低,且具有腫瘤微環(huán)境敏感性釋藥、經(jīng)濟(jì)性好等優(yōu)勢(shì),因此具有工業(yè)化、商品化的價(jià)值。體內(nèi)外研究結(jié)果表明,該載體材料能夠保護(hù)藥物在體內(nèi)循環(huán)時(shí)不被過早的降解,且將化療藥物高效地靶向遞送至腫瘤部位,且材料自身降解,保持低毒性。本課題的第一部分利用半胱氨酸催化激活硫辛酸五元環(huán)斷裂后相互交聯(lián),并對(duì)交聯(lián)合成條件進(jìn)行篩選,后采用超聲乳化法合成制備硫辛酸納米粒。對(duì)載體材料進(jìn)行紫外驗(yàn)證及GPC分子量檢測(cè)均證實(shí)合成成功。對(duì)納米粒進(jìn)行特征性評(píng)價(jià),結(jié)果顯示納米粒徑在110nm左右,電位約為-35mv,呈近似球形。納米粒穩(wěn)定性良好,且多西他賽載藥量達(dá)4.51%±0.49%,并具有腫瘤微環(huán)境敏感性釋藥特性。本課題的第二部分對(duì)硫辛酸納米粒進(jìn)行了體外細(xì)胞學(xué)評(píng)價(jià),以肺癌A549細(xì)胞為目標(biāo)細(xì)胞,結(jié)果顯示空白納米材料細(xì)胞毒性較低,細(xì)胞攝取率相比于單體明顯增強(qiáng),約為其10倍,且與PLGA納米粒相當(dāng),細(xì)胞攝取途徑主要為網(wǎng)格蛋白及質(zhì)膜微囊介導(dǎo)的內(nèi)吞作用,且細(xì)胞攝取定位表明,硫辛酸納米�？刹糠痔右萑苊阁w。CCK-8、細(xì)胞凋亡及周期研究結(jié)果均證實(shí)包載多西他賽的納米粒其體外腫瘤細(xì)胞殺傷性藥效明顯增強(qiáng),其IC50值相對(duì)于單體的277.84 ng/ml,降低至79.62 ng/ml,且相比于PLGA-DTX納米粒的135.61 ng/ml亦有明顯減少。本課題的第三部分對(duì)硫辛酸納米粒進(jìn)行了體內(nèi)研究。結(jié)果表明硫辛酸納米粒具有良好的腫瘤靶向性,其經(jīng)尾靜脈注射后能較好的在腫瘤部位發(fā)生蓄積,且相比于PLGA納米�？梢杂行У販p少脾臟、肺臟的毒性作用。體內(nèi)抗腫瘤藥效評(píng)價(jià)亦證實(shí)其抗腫瘤藥效明顯增強(qiáng),其腫瘤抑制率由單體的64.8%上升至81.62%,且相比于PLGA-DTX納米的76.96%亦有上升。
[Abstract]:Tumor nano-targeted chemotherapy (nanotarget chemotherapy) refers to the delivery of chemotherapeutic drugs to tumor sites using nanotechnology in order to maximize the efficacy and reduce the side effects of chemotherapeutic drugs. The key of nano-targeted tumor therapy lies in nano-carrier materials. Good nano-carriers can protect the drug from premature degradation in vivo, and release the drug smoothly after reaching the target site, and self-degradation metabolism as non-toxic metabolites. Therefore, the development of new good nano-drug carrier materials has some challenges, and has a very important significance. Based on the microenvironment of tumor cells and related studies, a novel biodegradable polymer carrier material, crosslinked lipoic acid, was synthesized. This material has the advantages of simple composition, simple and controllable preparation process, good stability, low toxicity, and has the advantages of sensitive release of tumor microenvironment, good economy and so on, so it has the value of industrialization and commercialization. The results in vivo and in vitro showed that the carrier material could protect the drug from premature degradation in vivo circulation, and deliver chemotherapeutic drugs to tumor site efficiently, and the material itself degraded and kept low toxicity. In the first part of this paper, the cysteine was used to catalyze the catalytic activation of the quaternary ring of lipoic acid and cross-linking each other, and the conditions of cross-linking synthesis were screened, and then the phacoemulsification method was used to synthesize the lipoic acid nanoparticles. UV verification and GPC molecular weight detection of the carrier material confirmed that the synthesis was successful. The characteristics of the nanoparticles were evaluated. The results showed that the particle size was about 110nm and the potential was about -35 MV, which was approximately spherical. The nanoparticles were stable, and the drug loading of docetaxel was 4.51% 鹵0.49, and the drug release was sensitive to tumor microenvironment. In the second part of this study, the in vitro cytological evaluation of lipoic acid nanoparticles was carried out. The results showed that the cell toxicity of the blank nano-materials was lower, and the uptake rate of the cells was about 10 times higher than that of the monomers. In the same way as PLGA nanoparticles, the main pathway of cell uptake was endocytosis mediated by grid protein and plasma membrane microencapsulation, and the location of cell uptake showed that, Lipoic acid nanoparticles could escape lysosome. CCK-8. The results of apoptosis and cell cycle study confirmed that doxetaxel nanoparticles could significantly enhance the killing effect of tumor cells in vitro. The IC50 value decreased to 79.62 ng/ml, relative to 277.84 ng/ml, of monomers and significantly decreased compared with 135.61 ng/ml of PLGA-DTX nanoparticles. In the third part of this paper, the in vivo study of lipoic acid nanoparticles was carried out. The results showed that lipoic acid nanoparticles had good tumor targeting ability and could accumulate well in tumor site after injection through tail vein. Compared with PLGA nanoparticles, it could effectively reduce the toxicity of spleen and lung. The evaluation of anti-tumor effect in vivo also confirmed that its anti-tumor effect was obviously enhanced, its tumor inhibition rate increased from 64.8% of monomer to 81.62%, and it also increased compared with 76.96% of PLGA-DTX nanoparticles.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R943;R96
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本文編號(hào)：2203480