新型光敏剂4SB的光谱特性及与血清蛋白相互作用的研究

目的研究新型光敏剂17,4-位氨基磺酸竹红菌乙素衍生物（17-4-amino-1-butane-sulfonic acid-hypocrellin B,4SB）的光谱特性及其与人血清白蛋白（human serum albumin,HSA）的相互作用规律,为4SB临床和基础研究提供实验依据。方法使用分光光度计和荧光光谱仪测量4SB在PBS和HSA溶液中的吸收光谱和荧光光谱,并与临床使用的光敏剂血卟啉衍生物（hematoporphyrin derivative,HpD）作对比,研究HSA对4SB光谱的影响。将4SB溶于HSA溶液后持续震荡,不同的时间点测定4SB吸收光谱和荧光光谱,观察4SB与HSA作用的时间规律。观察HSA对4SB吸收和荧光光谱的影响：将浓度为0、1.25、2.5、5.0、10和20μM/ml的HSA加入5μM/ml的4SB中,测定吸收光谱和荧光光谱。HSA与4SB相互的荧光淬灭：将不同浓度的4SB加入HSA中,测定4SB的荧光光谱;将不同浓度的4SB加入HSA中,测定HSA的荧光光谱。结果 4SB在600 nm以上的波段较HpD的吸收有很大提高,在人血清溶液体系中此峰红移且增强了2.3倍。4SB与HSA的结合时间是30 min。HSA与4SB是1对1的特异性结合,且位点在色氨酸残基附近。结论 4SB在红光波段吸收好,与白蛋白结合后增强了对光的吸收,与白蛋白1∶1结合,是一种有开发前景的光敏剂。

The Spectral Property of 4SB and Interaction between 4SB and HSA

Objective To study the spectral property of 17-4-amino-1-butane-sulfonic acid-hypocrellin B（4SB） and interaction between 4SB and human serum albumin（HSA）.Methods The absorption spectra and fluorescence spectra of 4SB at different times after mixture 4SB with HSA for observing the influ-ence of interaction time.For observing the influence of HSA on the absorption spectra and fluorescence spectra of 4SB,measured the spectra of 5 μM/ml 4SB interacting with difference concentration of 0,1.25,2.5,5,10,and 20 μM/ml HSA.For observing the fluorescence quenching between 4SB and HSA,measured the fluorescence of 4SB and HAS after mixing 10 μM/ml HSA with difference concentration of 4SB.Results The value of absorption of 4SB was higher than that of HpD at the range above 600 nm,and the peak moved to red range and increased 2.3 times when mixed with HSA.The time of completely reaction between 4SB and HSA was 30 min.The ratio of 4SB binding with HSA was 1：1,and binding site was near the tryptophan residues.Conclusions 4SB is a potential photosensitizer because its good absorption in red range which enhanced by binding with HSA in the ratio of 1：1.