苯肾上腺素荧光标记物的合成及其生物活性的鉴定

Synthesis of BODIPY-FL-Iabeled phenylephrine and the determination of its biological activity

目的：合成α_1-肾上腺素受体特异性激动剂——苯肾上腺素(phenylephrine，PE)荧光标记物，检测其药理学生物活性。方法：用化学合成的方法将绿色荧光染料 BODIPY-FL 和 PE 进行微量缩合反应，薄层色谱法分离和纯化，用薄层色谱和质谱分析对其纯度和分子结构进行鉴定。通过分子生物学 Western blot 方法检测其药理学生物活性。结果：将 BODIPY-FL 与 PE 在无水、无氧和避光条件下进行羧合反应，经薄层色谱分离、纯化得到具有荧光的新化合物——BODIPY-FL-PE，结构经紫外光谱和质谱分析得到证实。Western blot 实验表明 BODIPY-FL-PE与 PE 同样具有激动α_1-肾上腺素受体，使胞外信号调节激酶(extracellular signal-regulated kinase，ERK)激活的药理作用。结论：经微量缩合反应合成的 BODIPY-FL-PE 具有α_1-肾上腺素受体激动剂药理学特性，可以用于肾上腺素受体行为观测，为进一步研究活细胞中肾上腺素受体的动态行为提供了工具。

Objective:To'synthesize BODIPY-FL-labeled phenylephrine(BODIPY-FL-PE)and deter-mine its biological activity.Methods:Condensation of BODIPY-FL(green fluorescence dye)and phe-nylephrine(α_1-adrenoceptor agonist)was performed by adding dicyclohexylcarbodiimide(DCC)in thepresence of absolute tetrahydrofuran(THF).The reaction occurred in absolutely oxygen and water condi-tion at room temperature.The crude product was separated and purified by thin-layer chromatography(TLC).The structure of BODIPY-FL-PE was characterized by TLC and mass spectrometry(MS).Itspharmabiological activity was determined by Western blot.Results:BODIPY-FL-PE,the target mole-cule,was synthesized and its structure was identified by using ultra-violet spectrometry(UV)and MS.The result of Western blot indicated that α_1-adrenoceptor(α_1-AR)induced ERK phosphorylation wasconfirmed in both BODIPY-FL-PE and PE treated groups.Conclusion:The synthesized BODIPY-FL-PEhas pharmacological activity that could activate α_1-AR.Visualization of AR behaviors could be achievedby tracing the trajectories of BODIPY-FL-PE labeled AR.It might be a promising tool for investigatingdynamic behaviors of AR in living cells.