苯氧乙酸类化合物对人谷胱甘肽-S-转移酶Mu抑制性能表征  

Characterization of phenoxyacetic acid compounds in inhibition of human glutathione-S-transferase Mu

作　　者：许榜田 方大树 谢雨晴 杨晓兰[2] XU Bangtian;FANG Dashu;XIE Yuqing;YANG Xiaolan(Department of Pharmaceuties,University-Town Hospital of Chongqing Medical University,Chongqing,401331;Key Laboratory of Clinical Laboratory and Diagnostics of Ministry of Education,College of Laboratory Medicine,Chongqing Medical University,Chongqing,400016,China)

机构地区：[1]重庆医科大学附属大学城医院药学部,重庆401331 [2]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016

出　　处：《陆军军医大学学报》2022年第18期1826-1834,共9页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(31570862);重庆市自然科学基金资助项目(cstc2019jcyj-msxmX0166);重庆医科大学附属大学城医院科研青苗培育项目(2021JC03)。

摘　　要：目的 设计合成4个苯氧乙酸类化合物,通过不同方法表征其对人谷胱甘肽-S-转移酶Mu(glutathione-S-transferase Mu, GSTM)的抑制性能,考察苯氧乙酸类化合物与GSTM之间的构效关系。方法 设计合成含α,β-不饱和酮结构的4-丙烯酰苯氧乙酰乙胺(Ⅰa)、N,N’-(4-(丙烯酰基)苯氧乙酰基)丁二胺(Ⅱa)和不含α,β-不饱和酮结构的4-丙酰苯氧乙酰乙胺(Ⅰb)、N,N’-(4-(丙酰基)苯氧乙酰基)丁二胺(Ⅱb),利用初速度法表征4个化合物对GSTM的半抑制浓度(50%inhibiting concentration,IC)和表观抑制常数(inhibition constant,K),利用荧光光谱法表征4个化合物对GSTM的解离常数(dissociation constant,K)。结果 含α,β-不饱和酮结构的Ⅰa、Ⅱa可与谷胱甘肽(glutathione, GSH)发生亲电加成反应,与酶和GSH孵育原位生成产物后,其对GSTM的IC分别为(67±6)μmol/L、(0.10±0.02)μmol/L (n=2);不含α,β-不饱和酮结构的Ⅰb、Ⅱb对GSTM无明显抑制作用。Ⅰa相对GSH的K为(19.9±1.6)μmol/L,相对CDNB的K为(9.1±0.7)μmol/L (n=3);Ⅱa相对GSH的K为(0.063±0.005)μmol/L,相对CDNB的K为(0.079±0.006)μmol/L (n=3);可见Ⅱa较Ⅰa的K低(P<0.05)。荧光光谱法测得Ⅰa、Ⅰb、Ⅱa、Ⅱb的K分别为(41.5±1.8)μmol/L、(38.9±1.7)μmol/L、(18.0±0.9)μmol/L、(19.5±1.0)μmol (n=2);可见Ⅰa与Ⅰb、Ⅱa与Ⅱb之间的K无显著差异(P>0.05),而Ⅱa较Ⅰa、Ⅱb较Ⅰb的K低(P<0.05)。结论 α,β-不饱和酮结构是苯氧乙酸类化合物抑制作用的必须官能团;二价抑制剂Ⅱa、Ⅱb较单价抑制剂Ⅰa、Ⅰb对GSTM具有更高亲和力,Ⅱa的产物较Ⅰa的产物抑制作用更强。Objective To design and synthesize 4 phenoxyacetic acid compounds, characterize their inhibitory potencies on human glutathione-S-transferase Mu(glutathione-S-transferase Mu, GSTM) by different methods, and investigate the structure-activity relationship between phenoxyacetic acid compounds and GSTM. Methods Four phenoxyacetic acid compounds were designed and synthesized, and they are ethyl 2-(4-acryloylphenoxy)acetate(Ⅰa) and N,N’-(butane-1,4-diyl)-bis-(4-acryloyl phenoxyacetic amide)(Ⅱa) containing α,β-unsaturated ketone structure, and ethyl 2-(4-propionylphenoxy)acetate(Ⅰb), N,N’-(butane-1,4-diyl)-bis-(4-propionyl phenoxyacetic amide)(Ⅱb) not containing α,β-unsaturated ketone structure. They were characterized by the half-maximal inhibitory concentration(IC) and apparent inhibition constant(K) using initial velocity method, and the dissociation constants(K) for GSTM utilizing fluorescence analysis. Results Ⅰa and Ⅱa contained α,β-unsaturated ketone structure that undergoes electrophilic addition reaction with GSH. After incubation with enzyme and GSH, Ⅰa and Ⅱa generated products in situ, with a ICvalue of 67.00±6.00 and 0.10±0.02 μmol/L for GSTM respectively(n=2). Ⅰb and Ⅱb without α, β-unsaturated ketone structures had no inhibitory potencies for GSTM. The Kof Ⅰa against GSH was 19.9±1.6 μmol/L, and the Kagainst CDNB was 9.1±0.7 μmol/L(n=3). The Kof Ⅱa against GSH was 0.063±0.005 μmol/L, and against CDNB was 0.079±0.006 μmol/L(n=3). The Kof Ⅱa was lower than that ofⅠa(P<0.05). The Kof Ⅰa, Ⅰb, Ⅱa, and Ⅱb measured by fluorescence analysis were 41.5±1.8, 38.9±1.7, 18.0±0.9, and 19.5±1.0 μmol/L, respectively(n=2). The Kbetween Ⅰa and Ⅰb, and between Ⅱa and Ⅱb had no significant differences(P>0.05). The Kof Ⅱa was lower than that of Ⅰa,and the Kof Ⅱb was lower than that of Ⅰb(P<0.05). Conclusion α,β-unsaturated ketone structure is the essential functional group of phenoxyacetic acid compounds for inhibition on GSTM. Comparing with m

关 键 词：苯氧乙酸类化合物 谷胱甘肽-S-转移酶Mu 抑制性能 表征 

分 类 号：R345[医药卫生—基础医学] R914.4R962