Cytochrome P450-derived arachidonic acid metabolism in the rat kidney

Characterization of selective inhibitors

Mong Heng Wang, Elimor Brand-Schieber, Barbara A. Zand, Xuandai Nguyen, J R Falck, Narayanan Balu, Michal Laniado Schwartzman

Research output: Contribution to journalArticle

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Abstract

We characterized the inhibitory activity of several acetylenic and olefinic compounds on cytochrome P450 (CYP)-derived arachidonic acid ω- hydroxylation and epoxidation using rat renal cortical microsomes and recombinant CYP proteins. Among the acetylenic compounds, 6-(2- propargyloxyphenyl-)hexanoic acid (PPOH) and N-methylsulfonyl-6-(2- propargyloxyphenyl)hexanamide were found to be potent and selective inhibitors of microsomal epoxidation with IC50 values of 9 and 13 μM, respectively. On the other hand, 17-octadecynoic acid inhibited both ω- hydroxylation and epoxidation of arachidonic acid with IC50 values of 7 and 5 μM, respectively. The olefinic compounds N-methylsulfonyl-12,12- dibromododec-11-enamide (DDMS) and 12,12-dibromododec-11-enoic acid (DBDD) exhibited a high degree of selectivity inhibiting microsomal ω-hydroxylation with an IC50 value of 2 μM, whereas the IC50 values for epoxidation were 60 and 51 μM for DDMS and DBDD, respectively. Studies using recombinant rat CYP4A isoforms showed that PPOH caused a concentration-dependent inhibition of ω-hydroxylation and 11,12-epoxidation by CYP4A3 or CYP4A2 but had no effect on CYP4A1-catalyzed ω-hydroxylase activity. On the other hand, DDMS inhibited both CYP4A1- and CYP4A3- or CYP4A2-catalyzed arachidonic acid oxidations. Inhibition of microsomal activity by PPOH, but not DDMS, was time- and NADPH-dependent, a result characteristic of a mechanism-based irreversible inhibitor. These studies provide information useful for evaluating the role of the CYP-derived arachidonic acid metabolites in the regulation of renal function and blood pressure.

Original languageEnglish (US)
Pages (from-to)966-973
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume284
Issue number3
StatePublished - Mar 1998

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Arachidonic Acid
Cytochrome P-450 Enzyme System
Hydroxylation
Inhibitory Concentration 50
Kidney
Alkynes
Cytochrome P-450 CYP4A
Microsomes
Mixed Function Oxygenases
NADP
Protein Isoforms
DDMS
Blood Pressure
Acids
6-(2-propargyloxyphenyl)hexanoic acid
Proteins
cytochrome P-450 CYP4A2 (rat)

ASJC Scopus subject areas

  • Pharmacology

Cite this

Cytochrome P450-derived arachidonic acid metabolism in the rat kidney : Characterization of selective inhibitors. / Wang, Mong Heng; Brand-Schieber, Elimor; Zand, Barbara A.; Nguyen, Xuandai; Falck, J R; Balu, Narayanan; Schwartzman, Michal Laniado.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 284, No. 3, 03.1998, p. 966-973.

Research output: Contribution to journalArticle

Wang, MH, Brand-Schieber, E, Zand, BA, Nguyen, X, Falck, JR, Balu, N & Schwartzman, ML 1998, 'Cytochrome P450-derived arachidonic acid metabolism in the rat kidney: Characterization of selective inhibitors', Journal of Pharmacology and Experimental Therapeutics, vol. 284, no. 3, pp. 966-973.
Wang, Mong Heng ; Brand-Schieber, Elimor ; Zand, Barbara A. ; Nguyen, Xuandai ; Falck, J R ; Balu, Narayanan ; Schwartzman, Michal Laniado. / Cytochrome P450-derived arachidonic acid metabolism in the rat kidney : Characterization of selective inhibitors. In: Journal of Pharmacology and Experimental Therapeutics. 1998 ; Vol. 284, No. 3. pp. 966-973.
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abstract = "We characterized the inhibitory activity of several acetylenic and olefinic compounds on cytochrome P450 (CYP)-derived arachidonic acid ω- hydroxylation and epoxidation using rat renal cortical microsomes and recombinant CYP proteins. Among the acetylenic compounds, 6-(2- propargyloxyphenyl-)hexanoic acid (PPOH) and N-methylsulfonyl-6-(2- propargyloxyphenyl)hexanamide were found to be potent and selective inhibitors of microsomal epoxidation with IC50 values of 9 and 13 μM, respectively. On the other hand, 17-octadecynoic acid inhibited both ω- hydroxylation and epoxidation of arachidonic acid with IC50 values of 7 and 5 μM, respectively. The olefinic compounds N-methylsulfonyl-12,12- dibromododec-11-enamide (DDMS) and 12,12-dibromododec-11-enoic acid (DBDD) exhibited a high degree of selectivity inhibiting microsomal ω-hydroxylation with an IC50 value of 2 μM, whereas the IC50 values for epoxidation were 60 and 51 μM for DDMS and DBDD, respectively. Studies using recombinant rat CYP4A isoforms showed that PPOH caused a concentration-dependent inhibition of ω-hydroxylation and 11,12-epoxidation by CYP4A3 or CYP4A2 but had no effect on CYP4A1-catalyzed ω-hydroxylase activity. On the other hand, DDMS inhibited both CYP4A1- and CYP4A3- or CYP4A2-catalyzed arachidonic acid oxidations. Inhibition of microsomal activity by PPOH, but not DDMS, was time- and NADPH-dependent, a result characteristic of a mechanism-based irreversible inhibitor. These studies provide information useful for evaluating the role of the CYP-derived arachidonic acid metabolites in the regulation of renal function and blood pressure.",
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AU - Wang, Mong Heng

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AU - Zand, Barbara A.

AU - Nguyen, Xuandai

AU - Falck, J R

AU - Balu, Narayanan

AU - Schwartzman, Michal Laniado

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