磷酸膽堿化殼聚糖衍生物的合成、表征及生物學評價
發(fā)布時間：2018-08-14 11:57
【摘要】：本研究從細胞膜仿生的角度出發(fā)，設計合成一種新的殼聚糖磷�；苌铮粤柞０锋I的方式將細胞膜結(jié)構(gòu)單元磷酸膽堿基團偶聯(lián)到殼聚糖骨架上，研究其合成方法、生物學性能及其納米化方法。 為了合成磷酸膽堿化殼聚糖衍生物，我們研究比較了三種殼聚糖的均相磷酸膽堿化方法：(A)采用新型高極性六氟異丙醇作反應介質(zhì)，基于Antherton-Todd反應實現(xiàn)殼聚糖的直接磷酰化；(B)利用N-鄰苯二甲�；瘹ぞ厶菫橹虚g體，以二氯磷酰膽堿為磷酰化試劑；(C)利用6-O-三苯基甲醚化殼聚糖為中間體，基于Antherton-Todd反應實現(xiàn)磷�；�。研究表明合成路線(C)適合用來均相合成磷酸膽堿化殼聚糖衍生物。 應用合成路線(C)，，通過改變投料比，基于Antherton-Todd反應合成了三種不同取代度的水溶性磷酸膽堿化殼聚糖衍生物。NMR和FTIR譜圖上對應-N+(CH3)3基團吸收峰的出現(xiàn)表明磷酸膽堿基團成功偶聯(lián)到殼聚糖骨架的氨基上，根據(jù)1H NMR譜圖的峰強度比計算出三種殼聚糖衍生物的取代度分別為16%、27%和42%。GPC數(shù)據(jù)顯示，與殼聚糖相比，磷酸膽堿化殼聚糖衍生物的分子量有所降低，分子量分布有所拓寬；XRD、TGA、DSC和水溶性實驗表明，磷酸膽堿化殼聚糖衍生物的結(jié)晶性能和熱穩(wěn)定性均有不同程度地下降，但是其在水中的溶解性能得到了很大地提升，三種取代度的衍生物均可溶于pH=1-12的水溶液中。 細胞毒性實驗表明，3T3細胞與磷酸膽堿化殼聚糖衍生物共培養(yǎng)的相對增殖率在80%-110%之間，細胞毒性為0級或1級，屬于無細胞毒性范疇；血液相容性實驗表明，引入磷酸膽堿基團可以延緩衍生物的凝血時間，而且可以有效抑制血小板在其上的黏附與激活；與牛血清白蛋白(BSA)的相互作用表明，磷酸膽堿基團的引入可以有效抑制殼聚糖衍生物與BSA之間的相互作用，減小蛋白質(zhì)的構(gòu)象改變，這對避免激活因蛋白質(zhì)構(gòu)象改變而導致的不良生物反應具有重要意義。 磷酸膽堿化殼聚糖衍生物可自組裝形成納米粒子。研究表明，磷酸膽堿化殼聚糖衍生物仍可與三聚磷酸鈉進行離子交聯(lián)形成納米粒子，這些納米粒子呈現(xiàn)規(guī)則的球形結(jié)構(gòu)，粒徑在60-120nm之間，Zeta電位介于18-28mV；同時，磷酸膽堿化殼聚糖衍生物具有兩親性，可在中性水溶液中自組裝成具有疏水核親水殼的納米膠束，由低到高，三種取代度衍生物的臨界膠束濃度分別為0.129mg/mL，0.201mg/mL，0.256mg/mL。所形成的納米膠束粒徑范圍在70-110nm之間，Zeta電位接近于0，介于0-4mV之間。這兩類納米粒子有望應用于藥物/基因載體。
[Abstract]:In this study, a new phosphorylation derivative of chitosan was designed and synthesized from the point of view of cell membrane bionics. The phosphorylcholine group of cell membrane structure was coupled to the chitosan skeleton by phosphoramide bond, and the synthesis method of the derivative was studied. Biological properties and nanocrystalline methods. In order to synthesize chitosan derivative of choline phosphate, we studied and compared three homogenous choline phosphate methods: (A) used a new high polarity hexafluoroisopropanol as the reaction medium, and realized the direct phosphorylation of chitosan based on Antherton-Todd reaction; (B) used N-phthalic chitosan as intermediate and; (C) as phosphorylation reagent. 6-O- triphenylmethyl ether chitosan was used as intermediate to realize phosphorylation based on Antherton-Todd reaction. The results show that the synthetic route (C) is suitable for homogeneous synthesis of choline phosphate chitosan derivatives. By changing the feed ratio by using the synthetic route (C), Based on Antherton-Todd reaction, three kinds of water-soluble choline phosphate chitosan derivatives were synthesized. NMR and FTIR spectra showed that the corresponding absorption peaks of -N (CH3) _ 3 group were successfully coupled to the amino group of chitosan skeleton. According to the peak intensity ratio of 1H NMR spectrum, the substitution degree of the three chitosan derivatives is 16% and 42%.GPC data show that compared with chitosan, the molecular weight of chitosan derivative of choline phosphate is lower and the molecular weight distribution is wider. DSC and water solubility tests showed that the crystallization and thermal stability of choline phosphate chitosan derivatives decreased in varying degrees, but their solubility in water was greatly improved. All three derivatives of degree of substitution are soluble in the aqueous solution of pH=1-12. The results of cytotoxicity test showed that the relative proliferation rate was between 80% and 110%, and the cytotoxicity was grade 0 or grade 1, which belonged to the category of no cytotoxicity. The introduction of choline phosphate groups can delay the coagulation time of the derivatives and inhibit the adhesion and activation of platelets on them, and the interaction with bovine serum albumin (BSA) (BSA) suggests that, The introduction of choline phosphate group can effectively inhibit the interaction between chitosan derivatives and BSA and reduce the conformation change of protein, which is important for avoiding the activation of adverse biological reactions caused by protein conformation changes. Chitosan derivatives of choline phosphate can be self-assembled to form nanoparticles. The results show that the chitosan derivatives of choline phosphate can still be crosslinked with sodium tripolyphosphate to form nanoparticles. These nanoparticles have regular spherical structure and the potential of Zeta between 60-120nm is 18-28mV, at the same time, The chitosan derivatives of choline phosphate have amphiphilic properties and can self-assemble into nanomicelles with hydrophobic core hydrophilic shells in neutral aqueous solution. The critical micelle concentrations of the three derivatives are 0.129 mg / mL ~ 0.201 mg / mL ~ (-1) ~ 0.256 mg 路m ~ (-1) 路m ~ (-1), respectively, and the critical micelle concentrations of the three derivatives are 0.129 mg / mL ~ (-1) ~ 0.201 mg / mL ~ (-1) ~ 0.256 mg / m ~ (-1). The particle size range of the resulting micelles is close to 0 between 70-110nm and 0-4mV. These two kinds of nanoparticles are expected to be used in drug / gene carriers.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R318.08
本文編號：2182786
[Abstract]:In this study, a new phosphorylation derivative of chitosan was designed and synthesized from the point of view of cell membrane bionics. The phosphorylcholine group of cell membrane structure was coupled to the chitosan skeleton by phosphoramide bond, and the synthesis method of the derivative was studied. Biological properties and nanocrystalline methods. In order to synthesize chitosan derivative of choline phosphate, we studied and compared three homogenous choline phosphate methods: (A) used a new high polarity hexafluoroisopropanol as the reaction medium, and realized the direct phosphorylation of chitosan based on Antherton-Todd reaction; (B) used N-phthalic chitosan as intermediate and; (C) as phosphorylation reagent. 6-O- triphenylmethyl ether chitosan was used as intermediate to realize phosphorylation based on Antherton-Todd reaction. The results show that the synthetic route (C) is suitable for homogeneous synthesis of choline phosphate chitosan derivatives. By changing the feed ratio by using the synthetic route (C), Based on Antherton-Todd reaction, three kinds of water-soluble choline phosphate chitosan derivatives were synthesized. NMR and FTIR spectra showed that the corresponding absorption peaks of -N (CH3) _ 3 group were successfully coupled to the amino group of chitosan skeleton. According to the peak intensity ratio of 1H NMR spectrum, the substitution degree of the three chitosan derivatives is 16% and 42%.GPC data show that compared with chitosan, the molecular weight of chitosan derivative of choline phosphate is lower and the molecular weight distribution is wider. DSC and water solubility tests showed that the crystallization and thermal stability of choline phosphate chitosan derivatives decreased in varying degrees, but their solubility in water was greatly improved. All three derivatives of degree of substitution are soluble in the aqueous solution of pH=1-12. The results of cytotoxicity test showed that the relative proliferation rate was between 80% and 110%, and the cytotoxicity was grade 0 or grade 1, which belonged to the category of no cytotoxicity. The introduction of choline phosphate groups can delay the coagulation time of the derivatives and inhibit the adhesion and activation of platelets on them, and the interaction with bovine serum albumin (BSA) (BSA) suggests that, The introduction of choline phosphate group can effectively inhibit the interaction between chitosan derivatives and BSA and reduce the conformation change of protein, which is important for avoiding the activation of adverse biological reactions caused by protein conformation changes. Chitosan derivatives of choline phosphate can be self-assembled to form nanoparticles. The results show that the chitosan derivatives of choline phosphate can still be crosslinked with sodium tripolyphosphate to form nanoparticles. These nanoparticles have regular spherical structure and the potential of Zeta between 60-120nm is 18-28mV, at the same time, The chitosan derivatives of choline phosphate have amphiphilic properties and can self-assemble into nanomicelles with hydrophobic core hydrophilic shells in neutral aqueous solution. The critical micelle concentrations of the three derivatives are 0.129 mg / mL ~ 0.201 mg / mL ~ (-1) ~ 0.256 mg 路m ~ (-1) 路m ~ (-1), respectively, and the critical micelle concentrations of the three derivatives are 0.129 mg / mL ~ (-1) ~ 0.201 mg / mL ~ (-1) ~ 0.256 mg / m ~ (-1). The particle size range of the resulting micelles is close to 0 between 70-110nm and 0-4mV. These two kinds of nanoparticles are expected to be used in drug / gene carriers.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R318.08
【參考文獻】
相關期刊論文 前10條
1 常德富;王江濤;;殼聚糖的化學改性及其應用[J];日用化學工業(yè);2006年04期
2 李東旭;耿燕麗;;殼聚糖的改性及作為生物材料的應用研究[J];材料科學與工程學報;2006年02期
3 史宏燦,徐志飛,趙學維,秦雄,王文祖,孫康,高向陽;人工氣管材料體外細胞毒性和生物相容性研究[J];第二軍醫(yī)大學學報;2002年10期
4 吳清基,劉世英,張敏;甲殼質(zhì)縫合線的制備及研究[J];中國紡織大學學報;1998年05期
5 董炎明,吳玉松,王勉;甲殼素類液晶高分子的研究V.取代基個數(shù)及長度對羥乙基殼聚糖液晶性的綜合影響[J];高分子學報;2001年02期
6 張秀彩,鐘偉,孟晟,程為莊,杜強國;磷脂聚合物的研究進展[J];高分子通報;2004年01期
7 陳建勇,劉冠峰;殼聚糖改性及用其整理紡織品抗菌性能的研究[J];功能高分子學報;2002年02期
8 許晨,盧燦輝,丁馬太;殼聚糖季銨鹽的合成及結(jié)構(gòu)表征[J];功能高分子學報;1997年01期
9 劉慧娟;李鵬;張亞東;郭操;鄧uo元;蔡健煒;劉伯里;;葉酸與人血清白蛋白結(jié)合作用的光譜研究[J];光譜學與光譜分析;2009年07期
10 俞繼華,馮才旺,唐有根;甲殼素和殼聚糖的化學改性及其應用[J];廣西化工;1997年03期
本文編號：2182786
本文鏈接：http://www.wukwdryxk.cn/yixuelunwen/swyx/2182786.html
