[1]许榜田,方大树,谢雨晴,等.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[J].陆军军医大学学报(原第三军医大学学报),2022,44(18):1826-1834.
　XU Bangtian,FANG Dashu,XIE Yuqing,et al.Characterization of phenoxyacetic acid compounds in inhibition of human glutathione-S-transferase Mu [J].J Amry Med Univ (J Third Mil Med Univ),2022,44(18):1826-1834.
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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(/HTML )
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
44卷
期数:
2022年第18期
页码:
1826-1834
栏目:
基础医学
出版日期:
2022-09-30

文章信息/Info

Title:
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
重庆医科大学附属大学城医院药学部；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
 
Author(s):
XU Bangtian1;  FANG Dashu1;  XIE Yuqing1;  YANG Xiaolan2

1Department of Pharmaceutics, University-Town Hospital of Chongqing Medical University, Chongqing, 401331; 2Key Laboratory of Clinical Laboratory and Diagnostics of Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China

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
Keywords:
 ; 
分类号:
R345; R914.4; R962
文献标志码:
A
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, IC50)和表观抑制常数(inhibition constant, Ki)，利用荧光光谱法表征4个化合物对GSTM的解离常数(dissociation constant, Kd)。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IC50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Ki为(19.9±1.6) μmol/L，相对CDNB的Ki为(9.1±0.7) μmol/L (n=3)；Ⅱa相对GSH的Ki为(0.063±0.005) μmol/L，相对CDNB的Ki为(0.079±0.006) μmol/L (n=3)；可见Ⅱa较Ⅰa的Ki低(P<0.05)。荧光光谱法测得Ⅰa、Ⅰb、Ⅱa、Ⅱb的Kd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Kd无显著差异(P>0.05)，而Ⅱa较Ⅰa、Ⅱb较Ⅰb的Kd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。

Abstract:

ObjectiveTo 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. MethodsFour 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 (IC50) and apparent inhibition constant (Ki) using initial velocity method, and the dissociation constants (Kd) for GSTM utilizing fluorescence analysis. ResultsⅠa 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 IC50 value 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 Ki of Ⅰa against GSH was 19.9±1.6 μmol/L, and the Ki against CDNB was 9.1±0.7 μmol/L (n=3). The Ki of Ⅱa against GSH was 0.063±0.005 μmol/L, and against CDNB was 0.079±0.006 μmol/L (n=3). The Ki of Ⅱa was lower than that ofⅠa (P<0.05). The Kd of 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 Kd  between Ⅰa and Ⅰb, and between Ⅱa and Ⅱb had no significant differences(P>0.05). The Kd of Ⅱa was lower than  that of Ⅰa ,and the Kd of Ⅱb was lower than that of Ⅰb (P<0.05). Conclusionα,β-unsaturated ketone structure is the essential functional group of phenoxyacetic acid compounds for inhibition on GSTM. Comparing with monovalent Ⅰa and Ⅰb, divalent Ⅱa and Ⅱb have higher affinity for GSTM. The product of Ⅱa has inhibitory potency significantly higher than that of Ⅰa.

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更新日期/Last Update: 2022-09-23