光谱法研究苯烯莫德与血清白蛋白的作用机制  

Interaction mechanism research of Tapinarof and serum albumin by spectroscopy

作　　者：孙燕 严丽俊 张洁雪 王如意[2] 刘浩[2] SUN Yan;YAN Lijun;ZHANG Jiexue;WANG Ruyi;LIU Hao(Department of Pharmacy,the First Affiliated Hospital of the Naval Medical University,Shanghai200433,China;Department of Pharmacy,Guangdong Food and Drug Vocational College,Guangdong Province,Guangzhou510520,China)

机构地区：[1]海军军医大学第一附属医院药剂科,上海200433 [2]广东食品药品职业学院药学院,广东广州510520

出　　处：《中国医药导报》2025年第1期18-25,37,共9页China Medical Herald

基　　金：广东省医学科学基金项目(A2024054)。

摘　　要：目的探讨苯烯莫德分别与牛血清白蛋白(BSA)和人血清白蛋白(HSA)之间的相互作用并揭示其构象变化。方法运用荧光光谱、紫外光谱、同步荧光和三维荧光等,在分子水平上系统分析苯烯莫德与血清白蛋白(SA)的相互作用机制。利用Stern-Volmer方程式和Van’t Hoff方程式计算出苯烯莫德分别与BSA和HSA在298、303、310 K下的速率常数(Ksv)、猝灭速率常数(Kq)、结合位点数(n)、焓变(ΔH)、熵值(ΔS)和吉布斯自由能(ΔG)。结果随着苯烯莫德浓度的递增,BSA和HSA的紫外吸收光谱和荧光发射光谱均发生了变化,这两种SA的荧光强度明显降低,最大发射波长发生了峰位移,Ksv和Ka随温度升高而降低,n为1.0~1.5,ΔH、ΔS和ΔG均<0。同步荧光和三维荧乐的结果表明,BSA和HSA色氨酸(Trp)和酪氨酸(Tyr)残基与苯烯莫德生成了复合物,且苯烯莫德主要影响BSA和HSA中Trp残基的微环境,对Tyr残基的微环境影响较弱。此外,苯烯莫德与HSA的作用力明显强于BSA。结论苯烯莫德能够与SA生成复合物,导致SA的内源性荧光静态猝灭,苯烯莫德与SA主要以氢键和范德华力为作用力,反应为自发反应。苯烯莫德使用初始剂量应较大,与其他高结合率药物合用时,应考虑血药浓度增大的影响。Objective To investigate the interactions between Tapinarof and bovine serum albumin(BSA)and human serum albumin(HSA),and to reveal their conformational changes.Methods Fluorescence spectroscopy,ultraviolet spectroscopy,synchronous fluorescence,and three-dimensional fluorescence were used to systematically analyze the interaction mechanism between Tapinarof and serum albumin(SA)at the molecular level.Using the Stern Volmer equation and Van’t Hoff equation,the rate constant(Ksv),quenching rate constant(Kq),binding site number(n),enthalpy change(ΔH),entropy value(ΔS),and Gibbs free energy(ΔG)of Tapinarof with BSA and HSA were calculated at 298,303,and 310 K,respectively.Results As the concentration of Tapinarof increased,the UV absorption and fluorescence emission spectra of BSA and HSA changed.The fluorescence intensity of these two serum albumin(SA)decreased significantly,and the maximum emission wavelength shifted.Ksv and Ka decreased with increasing temperature,and then n was 1.0-1.5.ΔH,ΔS,andΔG were all<0.The results of synchronous fluorescence and three-dimensional fluorescence analysis indicated that BSA and HSA tryptophan(Trp)and tyrosine(Tyr)residues form complexes with Tapinarof,and Tapinarof mainly affected the microenvironment of Trp residues in BSA and HSA,with weaker effects on the microenvironment of Tyr residues.In addition,the interaction between Tapinarof and HSA was significantly stronger than that of BSA.Conclusion Tapinarof can form a complex with SA,leading to the static quenching of endogenous fluorescence of SA.Tapinarof and SA mainly react spontaneously through hydrogen bonding and van der Waals forces.The initial dose of Tapinarof should be relatively high,and the effect of increased blood drug concentration should be considered when used in combination with other high binding drugs.

关 键 词：光谱法 牛血清白蛋白 人血清白蛋白 苯烯莫德 

分 类 号：R96[医药卫生—药理学] O657.3[医药卫生—药学]