Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N′-(4-butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis
Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N′-(4-butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis
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DOI：
10.1248/bpb.28.1651
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发表时间：
2005-09-01
影响因子：
2
通讯作者：
Laniado-Schwartzman, M
中科院分区：
文献类型：
--
作者：
Seki, T;Wang, MH;Laniado-Schwartzman, M
我们研究了N-羟基-N '-（4-丁基-2-甲基苯基）-甲脒）（HET 0016），一种重组细胞色素P450 4A 1（CYP 4A 1）、CYP 4A 2和CYP 4A 3催化花生四烯酸（AA）ω-羟基化和环氧化的20-羟基-5，8，11，14-二十碳四烯酸（20-HETE）合成抑制剂，并表征了HET 0016的酶抑制特性。HET 0016对重组CYP 4A 1、CYP 4A 2和CYP 4A 3催化的20-HETE合成的IC 50值平均值分别为17.7 nM、12.1 nM和20.6 nM。CYP 4A 2和4A 3产生11，12-环氧-5，8，14-二十碳三烯酸（11，12-EET）的IC 50值平均值分别为12.7 nM和22.0 nM。CYP 2C 11活性的IC 50值为611 nM，远大于CYP 4A。初速度研究显示，HET 0016对CYP 4A 1的Ki值为19.5 nM，V-max与加入的重组CYP 4A 1量的关系图显示，HET 0016是一种不可逆的非竞争性抑制剂。这些结果表明，HET 0016是一种选择性、非竞争性和不可逆的CYP 4A抑制剂。
We examined the effect of N-hydroxy-N'-(4-butyl-2-methylphenyl)-formamidine) (HET0016), an inhibitor of 20-hydroxy-5,8,11,14-eicosatetraenoic acid (20-HETE) synthesis on the omega-hydroxylation and epoxidation of arachidonic acid (AA) catalyzed by recombinant cytochrome P450 4A1 (CYP4A1), CYP4A2 and CYP4A3, and characterized the enzyme inhibitory profile of HET0016. The IC50 values of HET0016 for recombinant CYP4A1-, CYP4A2- and CYP4A3-catalyzed 20-HETE synthesis averaged 17.7 nM, 12.1 nM and 20.6 nM, respectively. The IC50 value for production of 11,12-epoxy-5,8,14-eicosatrienoic acid (11,12-EET) by CYP4A2 and 4A3 averaged 12.7 nM and 22.0 nM, respectively. The IC50 value for CYP2C11 activity was 611 nM which was much greater than that for CYP4As. The initial velocity study showed the K-i value of HET0016 for CYP4A1 was 19.5 nM and a plot of V-max versus amount of recombinant CYP4A1 added shows HET0016 is an irreversible non-competitive inhibitor. These results indicate that HET0016 is a selective, non-competitive and irreversible inhibitor of CYP4A.