目的 构建简单有效的小分子药物长链生物素化探针合成方法。方法 采用一步法合成的生物素化氨基己酸作为中间体,以自发荧光的多柔比星∶(doxorubicin,DOX)作为实施例,经二环己基碳二亚胺(DCC)、4-二甲氨基吡啶(DMAP)催化,合成长链生物素化DOX;利用生物化DOX的双荧光系统,验证其亲和素结合能力、生物学活性。经验证后,应用于长链生物素化无荧光的紫杉醇(paclitaxel,PTX)的合成,并验证其理化活性、生物学活性及可视化效果。结果 该方法合成得到的生物素化DOX与亲和素结合率达93.7%;对细胞的抑制率及定位与DOX相同;生物素化PTX纯度为84.42%,核磁结构正确;对细胞的抑制效果同于PTX;通过可视化修饰可以观察到细胞内PTX与微管微丝的结合。结论 该合成方法可用于小分子药物生物素化探针的合成并具可视化用途,为药物机制研究提供了有效的工具。
Abstract
OBJECTIVE To set up an easy and effective method for biotinylation of small molecule drugs with long chain. METHODS Biotinylated 6-aminocaproic acid was synthesized as intermediate by one step method, doxorubicin(DOX) with auto-fluorescence was used as the first drug, and by DCC and DMAP catalysis, biotinylated DOX was synthesized. Using the double fluorescence system of DOX, the binding ability of biotinylated DOX to avidin and its biological activity were determined. When verified to be reasonable and effective, the method was applied to catalyze biotinylated paclitexal (PTX) which didn′t have auto-fluorescence itself, and the physical and chemical characteristics, and biological activities as well as the visualization were tested. RESULTS The binding rate of synthesized DOX to avidin was 93.7%; the cells inhibition rate and localization were the same as DOX; the purity of biotinylated PTX was 84.42%, and the structure shown by NMR was correct; the cell inhibition rate was the same as PTX; the combination of PTX with microtubules was observed by visual modification. CONCLUSION The method supplies a temperate way for biotinylation, and can be used for the synthesis and visualization of small molecules as probes and research of drug mechanism.
关键词
生物素 /
多柔比星∶ /
紫杉醇 /
6-氨基己酸 /
探针
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Key words
biotin /
doxorubicin /
paclitaxel /
6-aminocaproic acid /
probe
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中图分类号：
R914
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参考文献
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脚注
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基金
国家自然科学基金国际(地区)合作与交流项目资助(81361168001)
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