医用壳聚糖缓释材料研究
摘要
壳聚糖是自然界存在的唯一碱性多糖，无毒，具有体内生物降解性，壳聚糖是甲壳素通过强碱水解或酶解后脱去部分乙酰基的产物，是氨基葡萄糖的聚糖。壳聚糖相比甲壳素，通过化学处理已脱掉甲壳素中的乙酰基，可溶于稀酸，易被吸收利用，壳聚糖应用开发研究是当今国内外甲壳素类物质研究、开发的趋势。缓释制剂通过控制药物释放速率来获得所需的人体血药浓度，减少给药次数，降低人体不良反应程度，提高药效，是目前药物制剂开发方向。壳聚糖这类天然高分子用作缓释载体基材，相对人工合成材料，不仅可调控稳定的血药治疗浓度，减少给药次数，降低对胃肠道强烈刺激作用，且可参与人体自然代谢，在体内不会蓄积。
本课题研究被载药物选择盐酸氧氟沙星和水杨酸钠，前者是第三代氟喹诺酮类药物，对各种革兰氏阴性菌、阳性菌均有抗菌活性，半衰期6小时，而后者具有抗真菌、解热、镇痛、抗风湿作用，用于活动性风湿病及风湿性关节炎等的治疗。本研究以壳聚糖为载体主材，分别采用反相悬浮交联法，应用胺醛缩合、醇醛缩合原理和化学聚合技术，分别制备了壳聚糖载药缓释微球，壳聚糖和聚乙烯醇物理共混载药缓释微球、壳聚糖交联聚乙烯醇载药缓释微球、壳聚糖-聚丙烯酸-聚乙二醇三元复合缓释微球，所取得的主要研究内容和结果如下：
1、通过反相悬浮交联法制备了壳聚糖微球和盐酸左氧氟沙星载药微球，利用扫描电镜、显微镜、红外光谱、紫外分光光度仪和药物溶出仪对壳聚糖微球和载药微球进行表征，探讨了醋酸浓度、壳聚糖浓度、壳聚糖脱乙酰度、乳化剂用量、油水比、交联密度、反应时间、搅拌、药物／基材比等因素对微球平均粒径和粒径分散度、载药浓度、缓释性能的影响，可得到微球粒径，20～200μm，表面圆整、分散度可达0.3，盐酸左氧氟沙星壳聚糖微球载药量高达17％，12hr体外溶出累计释放度才达52％；水杨酸钠壳聚糖缓释微球载药量21.6％，释放曲线呈现出良好的线性关系，可持续释药8hr以上且具有优良的肠缓释给药特性。
2、以壳聚糖与聚乙烯醇醋酸溶液为水相，含乳化剂的液体石蜡为油相，分别采用室温改性方法(物理共混)制备载药复合微球Ⅰ和高温酸催化改性方法(化学交联)制备载药复合微球Ⅱ。通过红外光谱、显微镜、紫外分光光度仪和药物溶出仪等对材料结构和应用性能进行表征，结果表明壳聚糖／聚乙烯醇复合微球Ⅰ的平均粒径1～20μm，载药量13％，LVFX体外12hr累积释放80％；壳聚糖／聚乙烯醇复合微球Ⅱ的平均粒径1.69μm，载药量17.1％，LVFX体外6hr基本完全释放。
3、以含乳化剂的液体石蜡为油相，以壳聚糖／聚乙二醇醋酸溶液为水相，通过戊二醛使壳聚糖发生化学交联，反相悬浮聚合制备壳聚糖／聚乙二醇水杨酸钠载药微球。通过红外光谱、显微镜、紫外分光光度仪和药物溶出仪等对材料结构和应用性能进行表征，结果表明微球平均粒径60.0μm，分散度0.52，载药量达2.61％。
4、采用化学聚合和物理共混两种方法，制备壳聚糖／聚丙烯酸／聚乙二醇三元共混载SA微球。通过红外光谱、显微镜、紫外分光光度仪和药物溶出仪等对材料结构和应用性能进行表征，结果表明化学聚合方法制备微球平均粒径16.1μm，载药量达3.8％，物理共混方法制得微球粒径8.76μm，载药量达3.0％。
Chitosan is a kind of alkalescence amylase which is exclusive, nontoxic, and biodegradable. It is produced by part deacetylation of chitins using alkali or enzyme. Therefore, it can also be called as polysaccharide of amido glucose. After chitin is deacetylated by chemical method, chitosan is solve in weak acid, and its absorptivity is high. The application researches on chitosan are becoming its trend of research and development in the world.
Through controlling the drug release rate, the results of suitable drug concentration in the blood, less drug-delivery frequency, lower bad respondence, and increased curative effect have been achieced. This is the trend of research and development in drug manufacture recently. Compared with synthetic materials, chitosan using as the drug release carrier may not only adjust the drug concentration in blood, drug-delivery frequency, and drug stimulation on intestine and stomach, but also take part in the natural metabolism and accumulate nothing in the body.
In this article, chitosan is used as the main material. Levofloxacin and Sodium salicylate are choosen as the target drugs. Levofloxacin is the third generation of fluoroquinolones. Its half life is 6 hours, and its capbality of antibiotic is very good. Sodium salicylate has many advantages such as resisting fungus, curing fever, easing pain, and resisting rheumatism.
Using alcoholic aldehyde condensation reaction and reverse suspension-crosslink method, the chitosan drug-loaded microsphere, CS/PVA blending microsphere, CS/PAA/PEG complex microsphere were prepared. The brief contents and results are as follows:
1. Using reverse suspension-crosslink method, chitosan microsphere and chitosan drug-loaded microsphere were prepared. Those microspheres were characterized by microscope, IR spectrum, UV-4100, and drug dissolution test analyzer. The different influence factors on microsphere quality were discussed and the process was optimized. Finally, the uniform microspheres with good drug loading capability and drug releasing behavior are obtained.
2. Through polyer blending method and alcoholic aldehyde condensation reaction, PVA / chitosan microspheres with physical blending and chemical crosslinking structures are prepared. Their structures and properties were characterized by the microscope, IR spectrum, UV-4100, and drug dissolution test analyze. Finally, PVA / chitosan microspheres with physical blending are uniform microspheres about 1～20μm, drug loading capability to 13% and drug releasing percent to 80% in 12 hours. And PVA / chitosan microspheres with chemical crosslinking structures are uniform microspheres about 1.69μm, drug loading capability to 17.1% and drug releasing percent to 100% in 6 hours.
3. Using reverse suspension-crosslink method and physical blending method, CS/PEG complex microspheres were synthesized. Their structures and behaviours were characterized by the microscope, IR spectrum, UV-4100, and drug dissolution test analyzer. The process of CS/PEG microsphere was optimized. Finally, CS/PEG complex microspheres are uniform microspheres about 60μm and drug loading capability to 2.61%.
4. Using chemical polymerization and physical blending method, CS/PAA/PEG complex microspheres were synthesized. Their structures and behaviours were characterized by the microscope, IR spectrum, UV-4100, and drug dissolution test analyzer. The process of CS/PAA/PEG microsphere was optimized. Finally, CS/PAA/PEG complex microspheres with chemical polymerization are uniform microspheres about 16.1μm and drug loading capability to 3.8%.And CS/PAA/PEG complex microspheres with physical blending method are uniform microspheres about 8.76μm and drug loading capability to 3.0%.
本课题研究被载药物选择盐酸氧氟沙星和水杨酸钠，前者是第三代氟喹诺酮类药物，对各种革兰氏阴性菌、阳性菌均有抗菌活性，半衰期6小时，而后者具有抗真菌、解热、镇痛、抗风湿作用，用于活动性风湿病及风湿性关节炎等的治疗。本研究以壳聚糖为载体主材，分别采用反相悬浮交联法，应用胺醛缩合、醇醛缩合原理和化学聚合技术，分别制备了壳聚糖载药缓释微球，壳聚糖和聚乙烯醇物理共混载药缓释微球、壳聚糖交联聚乙烯醇载药缓释微球、壳聚糖-聚丙烯酸-聚乙二醇三元复合缓释微球，所取得的主要研究内容和结果如下：
1、通过反相悬浮交联法制备了壳聚糖微球和盐酸左氧氟沙星载药微球，利用扫描电镜、显微镜、红外光谱、紫外分光光度仪和药物溶出仪对壳聚糖微球和载药微球进行表征，探讨了醋酸浓度、壳聚糖浓度、壳聚糖脱乙酰度、乳化剂用量、油水比、交联密度、反应时间、搅拌、药物／基材比等因素对微球平均粒径和粒径分散度、载药浓度、缓释性能的影响，可得到微球粒径，20～200μm，表面圆整、分散度可达0.3，盐酸左氧氟沙星壳聚糖微球载药量高达17％，12hr体外溶出累计释放度才达52％；水杨酸钠壳聚糖缓释微球载药量21.6％，释放曲线呈现出良好的线性关系，可持续释药8hr以上且具有优良的肠缓释给药特性。
2、以壳聚糖与聚乙烯醇醋酸溶液为水相，含乳化剂的液体石蜡为油相，分别采用室温改性方法(物理共混)制备载药复合微球Ⅰ和高温酸催化改性方法(化学交联)制备载药复合微球Ⅱ。通过红外光谱、显微镜、紫外分光光度仪和药物溶出仪等对材料结构和应用性能进行表征，结果表明壳聚糖／聚乙烯醇复合微球Ⅰ的平均粒径1～20μm，载药量13％，LVFX体外12hr累积释放80％；壳聚糖／聚乙烯醇复合微球Ⅱ的平均粒径1.69μm，载药量17.1％，LVFX体外6hr基本完全释放。
3、以含乳化剂的液体石蜡为油相，以壳聚糖／聚乙二醇醋酸溶液为水相，通过戊二醛使壳聚糖发生化学交联，反相悬浮聚合制备壳聚糖／聚乙二醇水杨酸钠载药微球。通过红外光谱、显微镜、紫外分光光度仪和药物溶出仪等对材料结构和应用性能进行表征，结果表明微球平均粒径60.0μm，分散度0.52，载药量达2.61％。
4、采用化学聚合和物理共混两种方法，制备壳聚糖／聚丙烯酸／聚乙二醇三元共混载SA微球。通过红外光谱、显微镜、紫外分光光度仪和药物溶出仪等对材料结构和应用性能进行表征，结果表明化学聚合方法制备微球平均粒径16.1μm，载药量达3.8％，物理共混方法制得微球粒径8.76μm，载药量达3.0％。
Chitosan is a kind of alkalescence amylase which is exclusive, nontoxic, and biodegradable. It is produced by part deacetylation of chitins using alkali or enzyme. Therefore, it can also be called as polysaccharide of amido glucose. After chitin is deacetylated by chemical method, chitosan is solve in weak acid, and its absorptivity is high. The application researches on chitosan are becoming its trend of research and development in the world.
Through controlling the drug release rate, the results of suitable drug concentration in the blood, less drug-delivery frequency, lower bad respondence, and increased curative effect have been achieced. This is the trend of research and development in drug manufacture recently. Compared with synthetic materials, chitosan using as the drug release carrier may not only adjust the drug concentration in blood, drug-delivery frequency, and drug stimulation on intestine and stomach, but also take part in the natural metabolism and accumulate nothing in the body.
In this article, chitosan is used as the main material. Levofloxacin and Sodium salicylate are choosen as the target drugs. Levofloxacin is the third generation of fluoroquinolones. Its half life is 6 hours, and its capbality of antibiotic is very good. Sodium salicylate has many advantages such as resisting fungus, curing fever, easing pain, and resisting rheumatism.
Using alcoholic aldehyde condensation reaction and reverse suspension-crosslink method, the chitosan drug-loaded microsphere, CS/PVA blending microsphere, CS/PAA/PEG complex microsphere were prepared. The brief contents and results are as follows:
1. Using reverse suspension-crosslink method, chitosan microsphere and chitosan drug-loaded microsphere were prepared. Those microspheres were characterized by microscope, IR spectrum, UV-4100, and drug dissolution test analyzer. The different influence factors on microsphere quality were discussed and the process was optimized. Finally, the uniform microspheres with good drug loading capability and drug releasing behavior are obtained.
2. Through polyer blending method and alcoholic aldehyde condensation reaction, PVA / chitosan microspheres with physical blending and chemical crosslinking structures are prepared. Their structures and properties were characterized by the microscope, IR spectrum, UV-4100, and drug dissolution test analyze. Finally, PVA / chitosan microspheres with physical blending are uniform microspheres about 1～20μm, drug loading capability to 13% and drug releasing percent to 80% in 12 hours. And PVA / chitosan microspheres with chemical crosslinking structures are uniform microspheres about 1.69μm, drug loading capability to 17.1% and drug releasing percent to 100% in 6 hours.
3. Using reverse suspension-crosslink method and physical blending method, CS/PEG complex microspheres were synthesized. Their structures and behaviours were characterized by the microscope, IR spectrum, UV-4100, and drug dissolution test analyzer. The process of CS/PEG microsphere was optimized. Finally, CS/PEG complex microspheres are uniform microspheres about 60μm and drug loading capability to 2.61%.
4. Using chemical polymerization and physical blending method, CS/PAA/PEG complex microspheres were synthesized. Their structures and behaviours were characterized by the microscope, IR spectrum, UV-4100, and drug dissolution test analyzer. The process of CS/PAA/PEG microsphere was optimized. Finally, CS/PAA/PEG complex microspheres with chemical polymerization are uniform microspheres about 16.1μm and drug loading capability to 3.8%.And CS/PAA/PEG complex microspheres with physical blending method are uniform microspheres about 8.76μm and drug loading capability to 3.0%.
引文
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[68] 李药兰，黄才欢，岑颖洲，茶多酚缓释微囊的制备Ⅱ提高微囊药物包封率的研究，生物医学工程学杂志，1999，16(增刊)：115～116．
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[71] 孙伟张，曾仁杰，景利，等．喷雾干燥法制备阿司匹林肠溶微囊的实验研究，中国医院药学杂志，2000，20(7)：391～393．
[72] Genta-I. Different molecular weight chitosan microspheres: influence on drug loading and drug release. Drug Dev Ind Pharm, 1998, 24(8): 779～784.
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[74] Longo WE. Preparation of hydropholic albumin microspheres using polymeric dispersing agents [J], J. Pharm. Sci, 1982, 12: 1323.
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[82] 日本公开特许：昭61-268616[P]
[1] Lehr C, Bouwstra J. A., Schacht E. H., et al. In vitro evalution of mucoadhesive properties of chitosan and some other natural polymers. Int. J. Pharm., 1992, 78: 43.
[2] Tobio M., Gerf R., Sanchez A., et al. stealth PLA PEG nanoparticles as protein carriers for nasal administration. Pharm. Res., 1998, 15: 27.
[3] Giunchedi P., Genta I., Conti B., et al. Preparation and characterization of ampicillin loaded methylpyrrolidinone and chitosan microspheres. Biomaterials, 1998, 19: 157.
[4] Ramadas M., Paul W., Dileep K. J., et al. Liposulin encapsulated alginate- chitosan capsules intestinal delivery in diabetic rats. J. Microencapsulation, 2000, 17: 405.
[5] 王伟，薄淑琴，秦汶．中国科学，B辑．1990，(11)：1126．
[6] 蒋挺大．甲壳素(2003年第三版)，化学工业出版社，2003：70～71．
[7] LIU Li-Ping(刘利萍)，LI Ping(李苹)，WU Ze-Zhi(吴泽志)，et al．Chin Pharm J (中国药学杂志)[J]，2003，38(10)：775．
[8] Mima S. J. Appl Polym. Sci., 1983, 28: 1909.
[1] 邵强，张玉海，陈永和，等．左氧氟沙星治疗泌尿道感染的临床研究[J]．中国新药杂志，1999，8(1)：35～36．
[2] 中国药典(2000年版)二部．2000：447．
[1] R. A. A. Muzzarelli. Human enzymatic activities related to the therapeutic administration of chitin derivatives. New York: Plenum Publishing Co., 1990: 233.
[2] M. Rsukada, G.Freddi, J.Crightion. J. Polym. Sci.:Polym. Phys., 1994, 32: 243.
[3] 李沁华，邹翰．类肝素聚乙烯醇复合材料抗凝血性能的研究．中国生物医学工程学报，1999，18(1)：15．
[4] K. C. Gupta, M. N. V. Ravi Kumar. Semi-interpentrating polymer network beads of crosslinked chitosan-glycine for controlled release of chlorphertramine maleate. J. Appl. Polym. Sci., 2000,76: 672.
[5] Sangamessh G. Kumbar, Tejraj M. Aminabhavi. Synthesis and characterization of nifedipine through microspheres: J. Appl. Polym. Sci., 2003, 89: 2940.
[6] Xin-Yuan Shi, et al. Preparation of chitosan/ethylcellulose complex microcapsule and its application in controlled release of vitamin D_2. Biomaterials, 2002, 23: 4469.
[1] 代昭，孙多先，郭瑶．现代化工，2002，22(10)：22～25．
[2] K. C. Gupta, M. N. V. Ravi Kumar. Semi-interpentrating polymer network beads of crosslinked chitosan-glycine for controlled release of chlorphenramine maleate. J. Appl. Polym.Sci., 2000,76: 672.
[3] Sangamessh G.Kumbar, Tejraj M. Aminabhavi. Synthesis and characterization of nifedipine through microspheres: J. Appl. Polym. Sci., 2003, 89: 2940.
[4] Xin=Yuan Shi, Tian-Wei Tan. Preparation of chitosan/ethylcellulose complex microcapsule and its application in controlled release of vitarain D_2. Biomaterials, 2002, 23: 4469.
[5] 蒋新宇，周春山，张俊山．应用三聚磷酸钠为交联剂制备载药物纳米粒的研究，中国现代医学杂志，2003，13(22)：69～71．
[6] 大森英三著．丙烯酸酯及其聚合物．北京：化学工业出版社，1985．
[7] 蒋亚生．中国药学杂志，1987，22(10)：600．
[8] 刘砚星，李学配，刘俊秀，等．中国耳鼻咽喉头颈外科杂志，2005，40(11)：835～839．
[9] 王伟，薄淑琴，秦汶．中国科学，B辑．1990，(11)：1126．
[10] 蒋挺大．甲壳素(2003年第三版)，化学工业出版社，2003：70～71．
[1] 丁明，施建军，皇甫立霞，等．壳聚糖微球的制备研究，化学世界，1998，636～640．
[2] 钱国强，周菊岩，马建标，等．高等学校化学学报，1996，17(7)：1147．
[3] 中国药典(2000年版)二部．2000：447．