本文選題：肝靶向　切入點(diǎn)：羧甲基殼聚糖衍生物　出處：《山東大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：自組裝聚合物納米粒是由兩親性聚合物在水中通過疏水作用、靜電作用、氫鍵作用、金屬配位作用等驅(qū)動力,自組裝(或自聚集)形成具有親水性外殼和疏水性內(nèi)核的殼-核型納米微粒。自組裝聚合物納米粒具有較高的熱力學(xué)穩(wěn)定性,內(nèi)核可以作為疏水性藥物、基因、多肽和蛋白質(zhì)的貯庫,親水性外殼具有延長體內(nèi)循環(huán)時間的作用,并且可以通過化學(xué)修飾達(dá)到主動靶向、微環(huán)境響應(yīng)(pH敏感、溫度敏感、磁靶向)、逃避單核細(xì)胞的吞噬、提高生物膜轉(zhuǎn)運(yùn)等目的,因此自組裝聚合物納米粒在藥物遞送方面具有重大潛力,受到越來越多科研工作者的關(guān)注。 本文以水溶性O(shè)-羧甲基殼聚糖為骨架材料,對其進(jìn)行疏水性和靶向性修飾,用于構(gòu)造自組裝聚合物納米粒,作為抗腫瘤藥物的載體。首先,利用EDC催化的酰胺化反應(yīng)先后將具有疏水性烷基鏈的硬脂酸和具有肝靶向功能的乳糖酸接枝到O-羧甲基殼聚糖骨架上。利用紅外光譜法、核磁共振法以及元素分析法確證聚合物的結(jié)構(gòu)。采用超聲法制備自組裝納米粒并對其粒徑、表面電位進(jìn)行表征。由O-羧甲基殼聚糖接枝硬脂酸聚合物(OS)制備的納米粒粒徑在168～216nm范圍內(nèi),隨著硬脂酸取代度的增多,粒徑先減小后增大。選取硬脂酸取代度為9.8%的OS聚合物進(jìn)一步修飾半乳糖,得半乳糖化O-羧甲基殼聚糖接枝硬脂酸聚合物(Gal-OS),半乳糖取代度為13.1%。以此材料制備肝靶向自組裝聚合物納米粒,粒徑約為160nm。采用芘熒光探針法測量聚合物的臨界聚集濃度。隨著硬脂酸取代度的增加,OS聚合物CAC逐漸減小,而Gal-OS聚合物比具有相同硬脂酸取代度的未經(jīng)乳糖酸修飾的OS聚合物的CAC略大。溶血實(shí)驗(yàn)結(jié)果顯示,OS聚合物和Gal-OS聚合物的溶血率低于5%,說明材料血液相容性較好,符合靜脈注射的要求�？瞻撞牧系募�(xì)胞毒性實(shí)驗(yàn)表明,所制備的兩種材料對細(xì)胞沒有明顯的細(xì)胞毒性,可以應(yīng)用于生物醫(yī)學(xué)領(lǐng)域。將多柔比星作為治療藥物,制備Gal-OS/DOX自組裝納米粒。納米粒載藥量為13.4%,包封率為77.4%,粒徑為181.9nm。透射電鏡觀察納米粒形態(tài),為類球形實(shí)體,表面光滑、納米粒間不粘連。采用透析法考察Gal-OS/DOX和OS/DOX自組裝納米粒在三種不同pH值的釋放介質(zhì)中的釋放行為。結(jié)果表明,Gal-OS/DOX和OS/DOX納米粒體外釋放具有緩釋性,并且在較低pH的條件下釋藥更快,即具有pH敏感性。 本研究建立了熒光分光光度法測定多柔比星在血漿樣品和組織樣品中含量的分析方法。大鼠體內(nèi)藥動學(xué)研究結(jié)果顯示,Gal-OS/DOX和OS/DOX納米粒均能顯著延長多柔比星的體內(nèi)循環(huán)時間,降低清除速率。其藥-時曲線下面積分別為多柔比星溶液組的29倍和31倍。小鼠組織分布實(shí)驗(yàn)結(jié)果表明,Gal-OS/DOX和OS/DOX納米粒均能增加DOX在肝和脾的分布。其中,OS/DOX納米粒組在肝臟的AUC是溶液組的7.3倍,而Gal-OS/DOX納米粒組在肝臟的AUC是溶液組的9.8倍,表明Gal-OS/DOX納米粒比OS/DOX納米粒更能增加藥物在肝臟的積蓄。除此之外,兩種納米粒在心臟、腎臟以及肺的AUC比溶液組均要小,表明載藥納米粒均能降低藥物對心、腎以及肺的毒性,尤其是心臟毒性的降低,對多柔比星臨床應(yīng)用具有重大意義。靶向性評價結(jié)果說明,載藥納米粒對肝臟的趨向性大于多柔比星溶液組,且Gal-OS/DOX對肝臟的趨向性大于OS/DOX。 本文首次合成并報道了由乳糖酸、硬脂酸、O-羧甲基殼聚糖構(gòu)筑的新型肝靶向材料半乳糖化O-羧甲基殼聚糖接枝硬脂酸聚合物,并以此作為抗腫瘤藥物多柔比星的載體,制備肝靶向羧甲基殼聚糖衍生物自組裝納米粒制劑。本研究成果將為多柔比星的臨床應(yīng)用以及進(jìn)一步探討肝靶向給藥系統(tǒng)提供參考和新的選擇。
[Abstract]:The self-assembly of polymer nanoparticles is composed of two amphiphilic polymer in water by hydrophobic interaction, electrostatic interaction, hydrogen bonding, metal complexation, driving force, self assembly (or self aggregation) formed with hydrophilic shell and hydrophobic core shell nanoparticles. Karyotype thermodynamic stability of self-assembled polymer nanoparticles with high the kernel, can be used as hydrophobic drugs, gene, storage of peptides and proteins, the hydrophilic shell can prolong the circulation time in vivo, and can be reached by chemical modification of active targeting, response of micro environment (pH sensitive, temperature sensitive, magnetic targeting), phagocytosis of mononuclear cells escape, improve the biological membrane etc, so it has a great potential of self-assembled polymeric nanoparticles in drug delivery, more and more researchers' attention.
In this paper, the water soluble O- carboxymethyl chitosan as matrix material, the hydrophobic and targeted modification, for the construction of self assembled polymer nanoparticles as carrier of anti-tumor drugs. Firstly, stearic acid by amidation reaction catalyzed by EDC has a hydrophobic alkyl chain and lactose acid with liver graft targeting function to O- carboxymethyl chitosan skeleton. By infrared spectroscopy, nuclear magnetic resonance spectroscopy and elemental analysis confirmed the structure of the polymer. The ultrasonic preparation of self-assembled nanoparticles and their particle size and surface potential were characterized by O- carboxymethyl chitosan grafted stearic acid polymer (OS) prepared the particle size in the range of 168 ~ 216nm, with the degree of substitution of stearic acid increased, the particle size decreased and then increased. The degree of substitution of OS for selecting stearic acid polymer 9.8% galactose was further modified, galactosed O- carboxymethyl chitosan base Sugar grafted stearic acid polymer (Gal-OS), galactose substitution degree of 13.1%. taking the material preparation of liver targeted self-assembled polymeric nanoparticles, the particle size is about critical aggregation concentration of 160nm. using pyrene fluorescence probe method for the measurement of polymer. With the increase of degree of substitution of stearic acid, OS polymer CAC and Gal-OS gradually decreased, compared with the same polymer the degree of substitution of stearic acid without OS polymer lactose acid modified CAC slightly. The hemolysis experiment results show that OS polymer and polymer Gal-OS hemolytic rate of less than 5%, indicating the material has better blood compatibility, with intravenous injection requirements. Showed that the cytotoxicity of blank material, two kinds of materials prepared by no cell obvious toxicity to cells, can be used in the biomedical field. Doxorubicin as a treatment drug, preparation of Gal-OS/DOX nanoparticles self-assembled nanoparticles. The drug loading was 13.4%. The entrapment efficiency was 77.4%, grain The size of 181.9nm. transmission electron microscope to observe the morphology of nanoparticles were spherical, solid, smooth surface, no adhesion between the nanoparticles. The dialysis method was used to study the Gal-OS/DOX and OS/DOX self-assembled nanoparticles in three kinds of release medium of different pH value in the release behavior. The results showed that Gal-OS/DOX and OS/DOX nanoparticles in vitro release is sustained, and at low pH under the condition of drug release faster, which is the pH sensitivity.
This study established a method to measure than the stars in the plasma samples and tissue samples in soft fluorescence spectrophotometry. Pharmacokinetics in rats showed that Gal-OS/DOX and OS/DOX nanoparticles can significantly prolong the circulation time and more flexible than the stars, reduce the clearance rate. AUCs were doxorubicin solution were 29 times and 31 times. The tissue distribution in mice. The experimental results show that Gal-OS/DOX and OS/DOX nanoparticles can increase the distribution of DOX in the liver and spleen. Among them, OS/DOX nanoparticles group in the liver of the AUC is 7.3 times solution group, Gal-OS/DOX nanoparticles group in the liver of the AUC is 9.8 times solution group, showed that Gal-OS/DOX nanoparticles can increase the drug savings in the liver more than OS/DOX nanoparticles. In addition, two kinds of nanoparticles in the heart, kidney and lung of AUC group was smaller than that of solution, showed that the drug loaded nanoparticles can reduce the drug The toxicity of heart, kidney and lung, especially the decrease of cardiac toxicity, is of great significance for the clinical application of doxorubicin. The results of targeted evaluation indicate that the tendency of drug loaded nanoparticles to liver is greater than that of doxorubicin solution group, and the tendency of Gal-OS/DOX to liver is greater than OS/DOX..
This paper was first synthesized and reported by lactose acid, stearic acid, O- carboxymethyl chitosan to construct new liver targeting material galactosed O- carboxymethyl chitosan grafted stearic acid polymer, and as anticancer drug doxorubicin carrier, preparation of liver targeted carboxymethyl chitosan derivatives self-assembled nanoparticles. The results of this study will be to doxorubicin in clinical application and further study of liver target provide reference and new options to the drug delivery system.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R94

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 張闖年;王平;賴全勇;張倩;張s，

本文編號：1569174