两种组氨酸酰胺衍生物的合成及其与人血清白蛋白的结合

doi:10.11944/j.issn.1000-0518.2017.10.160473

摘要/Abstract

摘要：

L-组氨酸对生物有机体有着良好的亲和能力,通过修饰其化学结构以期寻找药理活性和生物利用度高的衍生物。本文将L-组氨酸分别与反式肉桂酸和对甲氧基肉桂酸反应,合成了两种组氨酸酰胺类衍生物,利用傅里叶变换红外光谱、质谱、氢谱/碳谱核磁共振谱进行了结构表征。采用分子操作环境(MOE)软件分子对接技术、荧光光谱法、同步荧光光谱法(SFS)、紫外-可见光谱法(UV-Vis),共同研究了两种衍生物分别和人血清白蛋白(HSA)相结合的机理。 MOE对接结果显示,这两种衍生物与HSA的模拟结合能分别为-13.82和-16.25 kcal/mol,主要是通过范德华力和疏水作用结合在HSA亚结构域ⅡA(即site Ⅰ)的疏水腔内。荧光猝灭数据表明,衍生物与HSA相互作用并形成了新的基态配合物,荧光猝灭过程为静态猝灭;不同温度(300、305和310 K)下衍生物与HSA相互作用的结合常数分别为1.773×10⁴、6.354×10³、1.260×10³和5.314×10⁴、4.614×10³、1.420×10³;由热力学参数得到衍生物与HSA的结合过程是由范德华力驱动;SFS表明,衍生物使得HSA的二级结构发生了变化。结合UV-Vis的结果可以确定,在体外生理条件下,组氨酸酰胺类衍生物均可以通过范德华力与HSA结合,并对HSA内源荧光产生静态猝灭及构象影响,这与分子对接结果一致,从而为组氨酸酰胺类衍生物药物的进一步开发提供了参考。

关键词: 组氨酸酰胺衍生物, 光谱法, 人血清白蛋白, 分子对接

Abstract:

L-Histidine has excellent affinity for biological organism. Its derivatives by structural modification may possess high pharmacological activity and bio-availability. In this work, two L-histidineamide derivatives were designed and synthesized by the reaction of L-histidine with trans-cinnamic acid and p-methoxycinnamic acid. Their structures were characterized by infrared, mass spectrometry, and nuclear magnetic resonance. The interaction mechanism of derivatives and human serum albumin(HSA) was investigated by molecular operating environment(MOE) molecular docking, fluorescence spectroscopy, synchronous fluorescence spectroscopy(SFS) and ultraviolet-visible(UV-Vis) absorption spectroscopy. The results of MOE molecular docking shows that the two derivatives exist in the hydrophobic pocket of subdomain ⅡA(site Ⅰ) of HSA under the action of van der Waals force and hydrophobic effect, with the simulation binding energy are -13.82 and -16.25 kcal/mol, respectively. The fluorescence quenching results show that the derivatives can interact with HSA and form new ground-state complexes and the fluorescence quenching process is a static quenching procedure. The binding of HSA to derivatives driven by van der Waals force was found from the thermodynamic parameters, and the binding equilibrium constants at different temperatures(300 K, 305 K, and 310 K) are 1.773×10⁴, 6.354×10³, 1.260×10³, 5.314×10⁴, 4.614×10³, 1.420×10³, respectively. The SFS characterization shows that the secondary structure of HSA has been changed by derivatives. Combining the results of UV-Vis spectra, it is obviously that under physiological conditions in vitro, the interaction between derivatives with HSA produces static quenching and conformational effects to the internal fluorescence of HSA through van der Waals force, which is consistent with the prediction of molecular docking, thus providing a reference for the further development of histidine amide derivatives research.

Key words: L-histidineamide derivatives, spectrometry methodology, human serum albumin, molecular docking

何蔚, 邹嘉佳, 逯东伟, 程辉, 林翠梧. 两种组氨酸酰胺衍生物的合成及其与人血清白蛋白的结合[J]. 应用化学, 2017, 34(10): 1150-1160.

HE Wei, ZOU Jiajia, LU Dongwei, CHENG Hui, LIN Cuiwu. Synthesis of Two L-Histidine Amide Derivatives and the Interaction Mechanism with Human Serum Albumin[J]. Chinese Journal of Applied Chemistry, 2017, 34(10): 1150-1160.

参考文献

[1]	YANG Feng,LIANG Hong.Structure of Human Serum Albumin and Its Complexes[J]. Prog Chem,2013,25(4):530-538(in Chinese). 杨峰,梁宏. 人血清白蛋白及其复合物的结构基础[J]. 化学进展,2013,25(4):530-538.
[2]	LIANG Jin,FENG Suling.Study on the Interaction Between Ambroxol Hydrochloride and Human Serum Albumin by Spectroscopy and Molecular Docking[J]. Spectrosc Spectr Anal,2011,31(4):1020-1024(in Chinese). 梁晶,冯素玲. 光谱法和分子对接法研究盐酸氨溴索与人血清白蛋白的相互作用[J]. 光谱学与光谱分析,2011,31(4):1020-1024.
[3]	Noguchi T,Inoue Y,Tang X S.Hydrogen Bonding Interaction Between the Primary Quinone Acceptor Qa and a Histidine Side Chain in Photosystem Ⅱ as Revealed by Fourier Transform Infrared Spectroscopy[J]. Biochemistry,1999,38(1):399-403.
[4]	Mathieu V,Van Den Berge E,Ceusters J,et al.New 5-Aryl-1H-Imidazoles Display in Vitro Antitumor Activity Against Apoptosis-Resistant Cancer Models, Including Melanomas, Through Mitochondrial Targeting[J]. J Med Chem,2013,56(17):6626-6637.
[5]	Petitpas I,Bhattacharya A A,Twine S,et al.Crystal Structure Analysis of Warfarin Binding to Human Serum Albumin Anatomy of Drug Site Ⅰ[J]. J Biol Chem,2001,276(25):22804-22809.
[6]	Kheder N A.Hydrazonoyl Chlorides as Precursors for Synthesis of Novel Bis-Pyrrole Derivatives[J]. Molecules,2016,21(3):326.
[7]	WANG Cunxin,CHANG Shan,GONG Xinqi,et al.Progress in the Scoring Functions of Protein-Protein Docking[J]. Acta Phys Chim Sin,2012,28(4):751-758(in Chinese). 王存新,常珊,龚新奇,等. 蛋白质—蛋白质分子对接中打分函数研究进展[J]. 物学学报,2012,28(4):751-758.
[8]	YU Yonghui.Study on Interactions and Recognition Between Ligand and Protein[D]. Beijing:Beijing University of Technology,2003(in Chinese). 于永辉. 配体—受体相互作用与识别的研究[D]. 北京:北京工业大学,2003.
[9]	Li D,Hong D,Guo H,et al.Probing the Influences of Urea on the Interaction of Sinomenine with Human Serum Albumin by Steady-State Fluorescence[J]. J Photochem Photobiol B,2012,117(117C):126-131.
[10]	Pacheco M E,Bruzzone L.Synchronous Fluorescence Spectrometry:Conformational Investigation or Inner Filter Effect[J]. J Lumin,2013,137(9):138-142.
[11]	Xie M X,Long M,Liu Y,et al.Characterization of the Interaction Between Human Serum Albumin and Morin[J]. Biochim Biophys Acta,2006,1760(8):1184-1191.
[12]	Yan Z Y,Lin X F,Yuzhu H U.Interaction Between Gatifloxacin and Bovine Serum Albumin[J]. J Chinese Pharm Sci,2005,14(1):33-37.
[13]	Ph.D. Haleem,J Issaq. Capillary Electrochromatography and Pressurized Flow Capillary Electrochromatography[J]. Instrum Sci Technol,2000,54(5):209-210.
[14]	Wang C,Wu Q H,Li C R,et al.Interaction of Tetrandrine with Human Serum Albumin:A Fluorescence Quenching Study[J]. Anal Sci,2007,23(4): 429.
[15]	Du C R,Luo X,Wei J R,et al.Preparation of (2E)-3-(4'-Halophenyl)prop-2-enoyl Sulfachlorpyridazine Sodium Salts and Their Interaction with Bovine Serum Albumin by Fluorescence Spectroscopy[J]. Chem Res Chinese Univ,2013,29(5):854-860.
[16]	ZHANG Qiong,SONG Yumin,LIU Jiacheng,et al.Interaction Between Complexes of Transition Metal with Warfarin and Human Serum Albumin[J]. Chinese J Inorg Chem,2011,27(9):1772-1780(in Chinese). 张琼,宋玉民,刘家成,等. 华法灵过渡金属配合物与人血清白蛋白相互作用研究[J]. 无机化学学报,2011,27(9): 1772-1780.
[17]	FENG Xiaoqiang,FU Guoqing,LI Xiaofang,et al.Interaction of Rare Earth with Bovine Serum Albumin[J]. Chinese J Lumin,2011,32(2):205-209(in Chinese). 冯小强,伏国庆,李小芳,等. 稀土La(Ⅲ)、Nd(Ⅲ)与牛血清白蛋白的相互作用[J]. 发光学报,2011,32(2):205-209.
[18]	ZHANG Yong,ZHANG Guizhu,WANG Yuemei,et al.Studies of Interaction of Mitomycin C and Serum Album in by Spectrum Method[J]. J Anal Sci,2000,16(6):445-449(in Chinese). 张勇,张贵珠,王月梅,等. 光谱法研究丝裂霉素、血清白蛋白以及金属离子间的相互作用[J]. 分析科学学报,2000,16(6):445-449.
[19]	WU Jinxiu,LI Mei,LIU Shaogang,et al.Study on Interaction of Y3+ and Bovine Serum Albumin by Spectrometry[J]. Chinese J Lumin,2012,33(10):1153-1159(in Chinese). 吴锦绣,李梅,柳召刚,等. 光谱法研究稀土离子钇(Ⅲ)与牛血清白蛋白的相互作用[J]. 发光学报,2012,33(10):1153-1159.
[20]	WANG Jian,ZHENG Xuefang,CAO Hongyu,et al.Synthesis of 4-Thio-5-iodouridine and the Interaction Between It and Human Serum Albumin[J]. Chinese J Lumin,2013,34(3):361-368(in Chinese). 王健,郑学仿,曹洪玉,等. 4-硫-5-碘尿苷的合成及其与人血清白蛋白间的相互作用[J]. 发光学报,2013,34(3):361-368.
[21]	Liu Y,Xie M X,Kang J,et al.Studies on the Interaction of Total Saponins of Panax Notoginseng and Human Albumin by Fourier Transform Infrared Spectroscopy[J]. Spectrochim Acta Part A,2003,59(12):2747-2758.
[22]	Mandal P,Bardhan M,Ganguly T.A Detailed Spectroscopic Study on the Interaction of Rhodamine 6g with Human Hemoglobin[J]. J Photochem Photobiol B,2010,99(2):78-86.
[23]	XU Li.Studies on Solution Thermodynamics of Protein and Its Model Molecules[D]. Zhengjiang University,2004(in Chinese). 许莉. 蛋白质模型分子溶液热力学研究[D].浙江大学,2004.
[24]	DU Chuanrong,LU Dongwei,SHI Kang,et al.Synthesis of Three Cinnamamide Derivatives and Their Interaction Mechanism with Human Serum Albumin[J]. Chinese J Lumin,2015,36(11):1342-1352(in Chinese). 杜传荣,逯东伟,石康,等. 三种肉桂酰胺衍生物的制备及其与人血清白蛋白的结合[J]. 发光学报,2015,36(11):1342-1352.