目的 构建低分子肝素pH敏感巯基壳聚糖纳米粒并对其进行表征;考察所构建纳米粒药物的肠黏膜透过作用。方法 采用离子交联法制备低分子肝素pH敏感巯基壳聚糖纳米粒，并对其体外特性进行表征;用Franz扩散池考察纳米粒的药物黏膜累积透过量。结果 纳米粒形态为类圆球形;平均粒径(329.6±7.3) nm，多相分散指数(0.185±0.01);电势(34.5±2.25) mV;包封率(97.75±0.02)%，载药量(12.463±0.003) u·mg-1;在pH 1.0介质中24 h累积释药:低分子肝素pH敏感巯基壳聚糖纳米粒＜巯基壳聚糖纳米粒＜药物溶液(P<0.05);在pH 6.8介质中24 h累积释药:pH敏感巯基壳聚糖纳米粒、巯基壳聚糖纳米粒均＜药物溶液(P<0.05);体外8 h药物肠黏膜累积透过量:pH敏感巯基壳聚糖纳米粒、巯基壳聚糖纳米粒＞pH敏感壳聚糖纳米粒＞药物溶液(P<0.05)，数值分别为(7.23±0.21) 、(6.99±0.79)、(5.62±0.22)、(3.88±0.76) u·cm-2。结论 所构建的低分子肝素pH敏感巯基壳聚糖纳米粒包封率高、粒径分布均匀，能增加低分子肝素肠黏膜透过率近1倍。
Abstract
OBJECTIVE To develop pH-sensitive thiolated chitosan nanoparticles and evaluate the mucosal permeation effect of the nanoparticles for low molecular weight heparin. METHODS The pH-sensitive thiolated chitosan nanoparticles were prepared by ionic cross-linking reaction and characterized for the shape, particle size, Zeta potential, drug entrapment efficiency and loading capacity. In vitro release of low molecular weight heparin from the prepared nanoparticles was evaluated in 0.1 mol稬-1 HCl solution and phosphate buffered saline (PBS) at pH 6.8 using the dialysis method. The intestinal permeability was estimated utilizing Franz diffusion cell system. RESULTS The obtained nanoparticles were found to be approximately spherical with the mean particles size of (329.6?7.3) nm and a positive Zeta potential of (34.5?2.25) mV; the entrapment efficiency and loading capacity of the NPs were (97.75?0.02)% and (12.463?0.003) u穖g-1, respectively. In 0.1 mol稬-1 HCl solution (pH 1.0) the release rate of low molecular weight heparin within 24 h from thiolated chitosan nanoparticles was faster than that from pH-sensitive thiolated chitosan nanoparticles with a significant difference (P<0.05).Whereas, in the case of release in PBS (pH 6.8), no noticeable difference was observed in the drug release behavior between pH-sensitive and non-pH-sensitive nanoparticles. The cumulative amount of permeability on carp intestine mucosa in vitro within 8 h were (7.23?0.21), (6.99?0.79), (5.62?0.22) and (3.88?0.76) u穋m-2 for pH-sensitive thiolated chitosan nanoparticles, thiolated chitosan nanoparticles, chitosan nanoparticles and low molecular weight heparin solution, respectively. CONCLUSION The pH-sensitive thiolated chitosan nanoparticles have a significant penetration enhancing effect for low molecule weight heparin.
关键词
低分子肝素 /
pH敏感 /
巯基壳聚糖 /
纳米粒 /
黏膜透过
{{custom_keyword}} /
Key words
low molecular weight heparin /
pH-sensitive /
thiolated chitosan /
nanoparticle /
mucosal penetration
{{custom_keyword}} /
中图分类号：
R944
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] LOIRA P C, SAPIN M A, DIAB R, et al. Low molecular weight heparin gels, based on nanoparticles, for topical delivery . Int J Pharm, 2012,426(1-2):256-262.[2] PALIWAL R, PALIWAL S R, AGRAWAL G P, et al. Chitosan nanoconstructs for improved oral delivery of low molecular weight heparin: In vitro and in vivo evaluation . Int J Pharm, 2012,422(1):179-184.[3] LEE C Y, LAI S T, SHIH C C, et al. Short-term results of catheter-directed intrathrombus thrombolysis versus anticoagulation in acute proximal deep vein thrombosis . J Chin Med Ass, 2013,76(5): 265-270.[4] HAYES P Y, ROSS B P, THOMAS B G, et al. Polycationic lipophilic-core dendrons as penetration enhancers for the oral administration of low molecular weight heparin . Bioorg Med Chem, 2006,14(1): 143-152.[5] JAVOT L, LECOMPTE T, RABISKOVA M, et al. Encapsulation of low molecular weight heparins: influence on the anti-Xa/anti-IIa ratio . J Controlled Release, 2009,139(1): 8-14.[6] OLIVEIRA S S M, OLIVEIRA F S, GAITANI C M, et al. Microparticles as a strategy for low-molecular-weight heparin delivery . J Pharm Sci, 2011,100(5): 1783-1792.[7] BAI S, AHSAN F. Synthesis and evaluation of pegylated dendrimeric nanocarrier for pulmonary delivery of low molecular weight heparin . Pharm Res, 2009,26(3): 539-548.[8] CHEN M C, WONG H S, LIN K J, et al. The characteristics, biodistribution and bioavailability of a chitosan-based nanoparticulate system for the oral delivery of heparin . Biomaterials, 2009,30(34): 6629-6637.[9] BAGRE A P, JAIN K, JAIN N K. Alginate coated chitosan core shell nanoparticles for oral delivery of enoxaparin: in vitro and in vivo assessment . Int J Pharm, 2013, 456(1): 31-40. FUENTES M G, ALONSO M J. Chitosan-based drug nanocarriers: Where do we stand . J Controlled Release, 2012,161(2):496-504. KAST C E, BERNKOP-SCHNRCH A. Thiolated polymers-thiomers: Development and in vitro evaluation of chitosan-thioglycolic acid conjugates . Biomaterials, 2001,22(17): 2345-2352. LIU D. Study on the preparation of a new type and ultra-pure low molecular weight heparin .Suzhou: Soochow University, 2010. YU S W,WANG F X,ZHAI G X, et al. Determination of low molecular weight heparinsodium by turbidimetric method .J Shandong Univ(山东大学学报), 2003,41(2):194-196. SHEN T. Studies on the oral drug delivery system of low molecular weight heparin-nanoemuision and intestinal adhesion stickers developed .Shanghai: Fudan University, 2005. LI F S, WANG Z P. Arsenic using cubic polynomial fitting spectrophotometric calibration curve . Nanjing Instit Chem Technol(南京化工学院学报),1989,11(2):38-40. GONG H M, FENG Z A, SHAO T J, et al. Encapsulation efficiency determination of low molecular weight heparin in isoliquiritigenin nanoparticle . Chin J Biochem Pharm (中国生化药物杂志), 2012,33(4):381-384. CHENA M C, MI F L, LIAO Z X, et al. Recent advances in chitosan-based nanoparticles for oral delivery of macromolecules . Adv Drug Deliv Rev, 2013,65(6):865-879. ANITHA A, DEEPA N, CHENNAZHI K P, et al. Development of mucoadhesive thiolated chitosan nanoparticles for biomedical applications . Carbohydrate Polymers, 2011,83(1): 66-73. ANDREAS B-S, MARGIT H, DAVIDE G. Thiolated chitosans . European J Pharm Biopharm, 2004,57(1):9-17. JAVED I, GUL S, SARAH D, et al. Preactivated thiomers as mucoad hesive polymers for drug delivery . Biomaterials, 2012,33(5):1528-1535. LI H H, HUANG P, DONG W, et al. A brief history of zebrafish research-toward biomedicine . Hereditas (遗传),2013, 35(4):410-420. JIA S J, MENG A M. The development of zebrafish research in China . Hereditas (遗传), 2012, 34(9):1082-1088. YA L, HUI D Z, MING K, et al. In vitro evaluation of mucoadhesion and permeation enhancement of polymeric amphiphilic nanoparticles . Carbohydrate Polymers, 2012,89(2):453-456.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
山西省自然科学基金资助项目(2014011047-4)；山西省大学生创新基金(2012130)
{{custom_fund}}
PDF(1547 KB)
