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Evaluation for adverse reaction of drugs using metabolic parameters

利用代谢参数评价药物不良反应

基本信息

批准号：
07672468
负责人：
HIRANO Kazuyuki
金额：
$ 1.41万
依托单位：
Gifu Pharmaceutical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672468/
关键词：
Nitroglycerine Glutathione S-tansferase Indomethacin Carboxylesterase Puranlukast Caco-2細胞エステラーゼ

项目摘要

Nitroglycerin (GTN) has been used as the drug of choice in the treatment of angina pectoris. It has been shown that some glutathione S-transferases (GSTs) catalyze the metabolic conversion from GTN to glyceryl dinitrates (GDNs). In this study, we examined the substrate specificity of GSTs for GTN.Alpha and mu GSTs were isolated from the liver and mucosa from pig, human and Caco-2 cells. Mu GSTs degraded GTN time-dependently and formed 1,3-GDN in preference to 1,2-GDN at ratio (1,2-GDN/1,3GDN) of 0.61, whereas alpha GSTs formed twice as much as different hydrolyzing portions of the nitrogroups.We found an enzyme involved in the hydrolysis of amide-linkage of indomethacin and partially characterized it as well as its substrate specificity. The purified enzyme effectively hydrolyzed the amide linkage in indomethacin but not those in a-naphthylacetate and p-nitrophenylacetate which are typical substrate for carboxylestrase. The Michaelis constant and maximum velocity values for indomethacin were 67 mM and 9 nmol/mg protein/min, respectively. This enzyme designated as ES-male. The activity of indomethacin hydrolysis was relatively high in the pig, rabbit and human liver, but not in those form rat and mouse. Two carboxylesteases with pl 6.0 and 6.2 derived from rat liver microsomes were purified. The two isozymes were remarkably different in substrate specificity. Carboxylesterases pl 6.0 and 6.2 are identical to the enzymes referred to as hydrolase A and B,respectively, from the results of amino acid sequence analyzes. Pranlukast was effectively hydrolyzed by caroxylesterase pl 6.2 but not by the pl 6.0 enzyme and the difference in the pranlukast metabolism between the human and the rat could be explained by the substrate specificity of carboxylesteases.

硝酸甘油（GTN）已被用作治疗心绞痛的首选药物。已经表明，一些谷胱甘肽S-转移酶（GST）催化从GTN到甘油二硝酸酯（GDN）的代谢转化。在这项研究中，我们研究了GST对GTN的底物特异性。从猪、人和Caco-2细胞的肝脏和粘膜中分离α和μ GST。Mu GSTs对GTN的降解具有时间依赖性，其1，2-GDN/1，3-GDN的比例为0.61，而α GSTs对不同硝基水解部位的GTN的生成量是其2倍。纯化的酶能有效水解吲哚美辛中的酰胺键，但不能水解羧基酯酶的典型底物α-萘乙酸酯和对硝基苯基乙酸酯中的酰胺键。吲哚美辛的米氏常数和最大速度值分别为67 mM和9 nmol/mg蛋白/min。这种酶被命名为ES-雄性。吲哚美辛在猪、兔和人肝中的水解活性较高，而在大鼠和小鼠肝中的水解活性较低。从大鼠肝微粒体中分离纯化了两种羧基酯酶，pI分别为6.0和6.2。两种同工酶的底物特异性差异显著。从氨基酸序列分析的结果来看，羧酸酯酶p16.0和6.2分别与称为水解酶A和B的酶相同。普仑司特可被胡萝卜素酯酶pl 6.2有效水解，但不能被pl 6.0酶水解，人和大鼠之间普仑司特代谢的差异可以通过羧酸酯酶的底物特异性来解释。