硬脂胺-异硫氰基荧光素嫁接物的合成工艺研究及其应用评价

Optimization of ODA-FITC Conjugate Process and Its Application

目的：探究硬脂胺-异硫氰基荧光素嫁接物的最优合成工艺,并对其进行应用评价。方法：以硬脂胺和异硫氰基荧光素为原料,通过偶合反应合成得到脂质材料硬脂胺-异硫氰基荧光素嫁接物,利用薄层色谱法监测反应过程;通过核磁共振对嫁接产物进行结构确证,并以溶剂、时间、温度以及投料比为考察因素,以产率为指标,得到最优合成工艺。并将合成的硬脂胺-异硫氰基荧光素嫁接物用于标记脂质纳米粒,以结扎肠循环模型考察纳米粒在小肠的吸收分布情况对其进行应用评价。结果：当反应溶剂为无水乙醇,硬脂胺和异硫氰基荧光素的反应摩尔比为2∶1,60℃搅拌反应48 h后异硫氰基荧光素与硬脂胺反应最完全,产率较高,可达（72.0±3.9）%,并能较准确地评价其标记的纳米粒在肠绒毛吸收分布情况。结论：优化后合成处方工艺反应完全,得率高;方法操作简单,重复性好;合成的硬脂胺-异硫氰基荧光素嫁接物可用于直观评价纳米粒的体内外情况。

Objective： To optimize the synthesis process of octadecylamine-fluorescein isothiocynate（ ODA-FITC）,and evaluate its application. Method： ODA-FITC was synthesized by coupling reaction with ODA and FITC as raw materials,and the reaction process was monitored by thin layer chromatography. Its structure was verified by nuclear magnetic resonance spectroscopy, and the optimum synthesis process was obtained with the solvent,time,temperature and feed ratio as the investigation factors and with yield as the indicator. ODA-FITC was then used as a fluorescence marker for nanostructured lipid carriers（ NLC） by solvent diffusion method. The ligated intestinal circulation model was established to observe the absorption and distribution of NLC in small intestine and evaluate its application. Result： The optimum reaction conditions were as follows： the molar ratio of ODA to FITC was 2 ∶ 1, the solution was anhydrous ethanol, after reaction for 48 h at 60 ℃. Under such conditions,the yield was high, reaching（ 72. 0 ± 3. 9） %, and the absorption and distribution of its marked nanoparticles in intestinal villus can be accurately evaluated. Conclusion： The synthesis method has complete reaction and high yield, with simple operation and high repeatability. Synthetic ODA-FITC can be used to intuitively evaluate the in vitro and in vivo conditions of nanoparticles.