Parij N, Nève J
Department of Organic Pharmaceutical Chemistry, Free University of Brussels, Belgium.
Eur J Pharmacol. 1996 Sep 12;311(2-3):259-64. doi: 10.1016/0014-2999(96)00427-x.
The established horseradish peroxidase/guaiacol in an in vitro assay system was used for investigation of the reactivity of nonsteroidal antiinflammatory drugs with hydrogen peroxide. Although the drugs rapidly seemed to react in the selected conditions, difficulties were encountered in attempts to quantify the reaction and an interaction with horseradish peroxidase was suspected. A more specific assay system based on the absolute specificity of the enzyme glutathione peroxidase for glutathione was subsequently used which demonstrated that none of the investigated nonsteroidal antiinflammatory drugs was able to scavenge hydrogen peroxide. An original procedure to further evidence the interaction was developed thereafter, based on the reaction of 5-aminosalicylic acid with similar hemoproteins. This led to the demonstration that nonsteroidal antiinflammatory drugs were substrates for horseradish peroxidase and explained their reactivity in the horseradish peroxidase/guaiacol assay system. The compound 5-aminosalicylic acid showed an unusual behaviour that was attributed to its ability to both scavenge hydrogen peroxide and interact with horseradish peroxidase. It was concluded that the lack of specificity of horseradish peroxidase for its donor substrate may lead to erroneous results in assays for hydrogen peroxide scavenging of some drugs. An alternative method is however available and a simple spectroscopic assay can evidence the interaction with horseradish peroxidase.
在体外分析系统中，使用已建立的辣根过氧化物酶/愈创木酚来研究非甾体抗炎药与过氧化氢的反应活性。尽管这些药物在选定条件下似乎迅速发生反应，但在试图对反应进行定量时遇到了困难，并且怀疑存在与辣根过氧化物酶的相互作用。随后使用了一种基于谷胱甘肽过氧化物酶对谷胱甘肽的绝对特异性的更特异的分析系统，该系统表明所研究的非甾体抗炎药均不能清除过氧化氢。此后，基于5-氨基水杨酸与类似血红蛋白的反应，开发了一种进一步证明相互作用的原始方法。这导致证明非甾体抗炎药是辣根过氧化物酶的底物，并解释了它们在辣根过氧化物酶/愈创木酚分析系统中的反应活性。化合物5-氨基水杨酸表现出异常行为，这归因于其清除过氧化氢和与辣根过氧化物酶相互作用的能力。得出的结论是，辣根过氧化物酶对其供体底物缺乏特异性可能导致在某些药物过氧化氢清除测定中产生错误结果。然而，有一种替代方法可用，一种简单的光谱分析可以证明与辣根过氧化物酶的相互作用。
