[关键词]
[摘要]
目的 在非水相中利用酶法合成脑靶向脂质材料胆固醇-癸二酸-葡萄糖酯[(5-cholesten-3β-yl)(D-glucopyranose-6)sebacate,CHS-SE-GLU],并对其修饰的载紫杉醇脂质体制备处方及其工艺进行优化。方法 以D-葡萄糖和前期合成的胆固醇-癸二酸单烯酯(CHS-SE)为原料,在丙酮中用利用脂肪酶Novozym 435催化合成CHS-SE-GLU,利用MS、NMR对产物进行结构表征,采用薄膜分散法制备CHS-SE-GLU修饰的脑靶向载紫杉醇脂质体(GLU-PTX-LP),并通过单因素考察对其制备处方及工艺进行优化。结果 合成的CHS-SE-GLU经MS、NMR鉴定为目标产物,对其修饰的GLU-PTX-LP最佳制备处方工艺为采用氢化大豆磷脂(HSPC)作为膜材,HSPC与紫杉醇(PTX)比例为0.1:1,胆固醇(CHS)与HSPC比例为0.5:1,二硬脂酰磷脂酰甘油钠(DSPG-Na)用量为2.5%,水化时间为0.5 h,水化温度为50℃。采用上述工艺处方制备3批样品,测定包封率分别为(93.62±1.34)%、(93.75±1.77)%、(92.04±1.50)%,平均粒径分别为(89.56±1.35)、(92.05±3.42)、(104.91±3.71)nm,Zeta电位分别为(-25.21±0.27)、(-26.43±0.44)、(-25.17±0.65)mV。结论 采用生物酶催化法合成脑靶向脂质材料,工艺简单,产率高,绿色环保;用其修饰的GLU-PTX-LP包封率、粒径及稳定性均符合要求,应用前景良好。
[Key word]
[Abstract]
Objective To synthesize brain targeting lipid material[(5-cholesten-3β-yl)(D-glucopyranose-6) sebacate, CHS-SE-GLU] by lipase as catalyst in nonaqueous phase and optimize the preparation technology and formulation of CHS-SE-GLU-modified liposomes. Methods CHS-SE-GLU was synthesized from CHS-SE prepared in previous work and D-glucose using lipase Novozym 435 in acetone. The structure characterization of the products is obtained by MS and NMR. The CHS-SE-GLU-modified paclitaxelloaded brain targeting liposomes (GLU-PTX-LP) were prepared by thin film dispersion method. Single factor evaluation was applied to optimizing its preparation technology and formulation. Results CHS-SE-GLU was confirmed by MS and NMR as target products. The optimal formulation and technology of GLU-PTX-LP were as follows:HSPC as membrane material, the ratio of HSPC to PTX was 0.1, the ratio of CHS to HSPC was 0.5, the dosage of DSPG-Na was 2.5%, hydration time was 0.5 h, and hydration temperature was 50℃. Three batches of samples were prepared by optimum preparation process and the average encapsulation efficiency was (93.62±1.34)%, (93.75±1.77)%, (92.04±1.50)%; The average particle size was (89.56±1.35), (92.05±3.42), (104.91±3.71) nm; And the average Zeta potential was (-25.21±0.27), (-26.43±0.44), (-25.17±0.65) mV, respectively. Conclusion Lipase-catalyzed method for the preparation of brain targeting lipid material is simple and environment friendly with high yield. The entrapment efficiency, particle size, and stability of brain targeting drug-loading liposomes modified by CHS-SE-GLU all meet the requirement, which shows good application prospect.
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[基金项目]
广东省自然科学基金项目(2016A030310365);广东省医学科研基金项目(A2015629);广东省省级科技计划项目(2014A020221061);梅州市科技研究与开发资金计划项目(201412);嘉应学院自然科学项目(2015KJY10)