Abstract: The natural macromolecular materials of pullulan(Pu)and hyaluronic acid(HA)have good biocompatibility and are often used as drug carriers. However, the rapid degradation of these natural macromolecule limits their application as drug sustained-release functional carriers. In this paper, the self-made anti-enzymatic degradation material of hyaluronic acid grafted pullulan(HA-Pu)is used to prepare its microspheres. In this system, HA-Pu material solution is used as the water phase, liquid paraffin as the oil phase, the Span 80 as the emulsifier, and the glutaraldehyde as the crosslinking agent to prepare the HA-Pu MPs. Box-Behnken Design(BBD)method was used to investigate the effects of rotational speed, oil-water ratio and HA-Pu concentration on the particle size of HA-Pu microspheres. When the optimal formulation conditions was selected as 670 r/min stirring speed, 5.6∶1(V/V, mL/mL)oil-water ratio and 4.48 g/mL HA-Pu, the microparticles had a smooth surface with about 18 μm diameter. The infrared spectrum- DOI:10.3969/j.issn.1000-1565.2020.01.006透明质酸基普鲁兰糖载药微球制备与体内外评价杨文智,赵亚非,吴桐,刘娇艳,李海鹰(河北大学 药学院,河北 保定 071002)摘 要:普鲁兰糖(Pu)和透明质酸(HA)天然高分子材料具备良好生物相容性,常用作药物载体,但此类天然高分子降解快,限制其作为药物的缓释功能载体的应用.本文拟采用自制抗酶降解的透明质酸接枝普鲁兰糖(HA-Pu)材料溶液为水相,液体石蜡为油相,Span 80为乳化剂,戊二醛为交联剂,利用乳化交联法制备HA-Pu微球(HA-Pu MPs). 利用Box-Behnken Design(BBD)法,考察转速、油水比和HA-Pu质量浓度等3因素对微球粒径的影响,当选择670 r/min搅拌转速,5.6∶1(体积比)油水比和44.8 mg/mL HA-Pu的最佳制备条件,可获得形态圆整且平均粒径约为18 μm的微球. 样品红外图谱显示,制备微球成功交联. 以阿霉素(DOX)为模型药,交联量戊二醛0.5 mol,最佳DOX与HA-Pu投料比为2∶10(质量比),获得载药量为5.02%(质量分数),包封率为33%的(载药微球)DOX-HA-Pu MPs. 载药微球体外释药曲线拟合符合Ritger-Peppas方程. 而对比大鼠尾静脉注射DOX药物和腹腔注射DOX-HA-Pu MPs,载药微球具备缓释功能. 利用HA-Pu材料抗酶降解性质,采用乳化交联法制备HA-Pu MPs,方法简单易行,制备微球有望成为抗瘤药物的缓释载体.关键词:微球;透明质酸;普鲁兰糖;盐酸阿霉素;体内外中图分类号:O63 文献标志码:A 文章编号:1000-1565(2020)01-0033-08Preparation and evaluation of doxorubicin loaded hyaluronic acid-pullulan microspheres in vitro and in vivo YANG Wenzhi, ZHAO Yafei, WU Tong, LIU Jiaoyan, LI Haiying(College of Pharmacy,Hebei University,Baoding 071002,China)Abstract: The natural macromolecular materials of pullulan(Pu)and hyaluronic acid(HA)have good biocompatibility and are often used as drug carriers. However, the rapid degradation of these natural macromolecule limits their application as drug sustained-release functional carriers. In this paper, the self-made anti-enzymatic degradation material of hyaluronic acid grafted pullulan(HA-Pu)is used to prepare its microspheres. In this system, HA-Pu material solution is used as the water phase, liquid paraffin as the oil phase, the Span 80 as the emulsifier, and the glutaraldehyde as the crosslinking agent to prepare the HA-Pu MPs. Box-Behnken Design(BBD)method was used to investigate the effects of rotational speed, oil-water ratio and HA-Pu concentration on the particle size of HA-Pu microspheres. When the optimal formulation conditions was selected as 670 r/min stirring speed, 5.6∶1(V/V, mL/mL)oil-water ratio and 4.48 g/mL HA-Pu, the microparticles had a smooth surface with about 18 μm diameter. The infrared spectrum- 收稿日期:2018-10-20 基金项目:河北省自然科学基金资助项目(H2017201052;H2018201045);河北大学研究生创新项目(hbu2019ss034);河北省高等学校科学技术项目(ZD2016102;ZD2018054) 第一作者:杨文智(1972—),男,内蒙古锡林浩特人,河北大学副教授,从事生物医用材料及药物缓控释制剂方面研究.第1期杨文智等:透明质酸基普鲁兰糖载药微球制备与体内外评价of the sample shows that the prepared microspheres are successfully crosslinked. By using doxorubicin(DOX)as a model drug, with the cross-linking amount of glutaraldehyde being 0.5 mol, and with the optimum drug loading mass ratio being 2∶10, DOX-HA-Pu MPs with 5% drug loading and 33% encapsulation efficiency were obtained. In vitro release curve of drug-loaded microspheres was consistent with the Ritger-Peppas equation. when DOX drugs were injected into tail vein of rats and DOX-HA-Pu MPs were injected intraperitoneally in rats, the drug-loaded microspheres have displayed a sustained-release function. In summary utilizing HA-Pu material anti-enzymatic degradation properties, HA-Pu MPs are prepared by emulsion cross-linking method which is simple and easy to operate. Therefore, the preparation of HA-Pu MPs is expected to be a sustained-release carrier of antineoplastic drugs.

Key words: microspheres, hyaluronic acid, pullulan, doxorubicin hydrochloride, in vitro and in vivo