Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N′-(4- butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis

Takayuki Seki, Mong-Heng Wang, Noriyuki Miyata, Michal Laniado-Schwartzman

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

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 ω-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 Ki 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.

Original languageEnglish (US)
Pages (from-to)1651-1654
Number of pages4
JournalBiological and Pharmaceutical Bulletin
Volume28
Issue number9
DOIs
StatePublished - Sep 1 2005

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Cytochrome P-450 Enzyme System
Protein Isoforms
Cytochrome P-450 CYP4A
Inhibitory Concentration 50
Hydroxylation
Arachidonic Acid
20-hydroxy-5,8,11,14-eicosatetraenoic acid
N-hydroxy-N'-(4-butyl-2-methylphenyl)formamidine
Enzymes
cytochrome P-450 CYP4A2 (rat)
11,12-epoxy-5,8,14-eicosatrienoic acid

Keywords

  • 20-hydroxy-5,8,11,14-eicosatetraenoic acid
  • Cytochrome P450 A4
  • HET0016
  • Kinetics

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Cite this

Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N′-(4- butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis. / Seki, Takayuki; Wang, Mong-Heng; Miyata, Noriyuki; Laniado-Schwartzman, Michal.

In: Biological and Pharmaceutical Bulletin, Vol. 28, No. 9, 01.09.2005, p. 1651-1654.

Research output: Contribution to journalArticle

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abstract = "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 ω-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 Ki 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.",
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AB - 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 ω-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 Ki 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.

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