Identification and characterization of human cytchrome P450 (CYP) isoforms interacting with cisapride

Zeruesenay Desta, David Thacker, Nadia Soukhova, Jae Gook Shin, David A. Flockhart

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Using human liver microsomes (HLMs) and recombinant CYP450s, we characterized the metabolism of cisapride and documented its inhibitory effect on CYP450s. Two cisapride enantiomers (EI and EII) were separated and fractions collected for study. HPLC chromatograms of HLM incubates of cisapride separated 3 metabolite peaks, M1-M3. The identity of M1 is consistent with norcisapride, while M2 and M3 may represent oxidative hydroxylation of the fluorophenyl ring. The formation rate of M1 and 10μM cisapride in 15 HLMs significantly correlated with the activities of CYP3A, 2C19 and 1A2. Of CYP isoform specific inhibitors tested, ketoconazole potently inhibited the formation of M1, M2 and M3. Of the 10 recombinant CYP450s tested, CYP3A4, 2C8, 2C19, 2C9, 1A2 and 2B6 formed M1. Inhibition by racemic cisapride of CYP2D6 was relatively potent (Ki=14±16μM), of CYP1A2 (Ki=40±12μM) moderate and of CYP3A (Ki=142±27μM) weak. The inhibition by cisapride of CYP2D6 appears to be due to EII as the IC50 (μM) of EII (20±6) was closer to that of racemic cisapride (10±1.8), while that of EI (59±17) was 6-fold higher. In conclusion, cisapride is metabolized by multiple CYP450s in vitro which may make prediction of cisapride drug interaction and cardiac toxicity more difficult than described in the manufacturer's labels. Cisapride appears to inhibit CYP2D6-mediated metabolism in a stereoselective manner.

Original languageEnglish (US)
Pages (from-to)126
Number of pages1
JournalClinical Pharmacology and Therapeutics
Volume65
Issue number2
StatePublished - 1999
Externally publishedYes

Fingerprint

Cisapride
Forensic Anthropology
Protein Isoforms
Cytochrome P-450 CYP3A
Cytochrome P-450 CYP2D6
Liver Microsomes
Cytochrome P-450 CYP1A2
Ketoconazole
Hydroxylation
Drug-Related Side Effects and Adverse Reactions
Drug Interactions
Inhibitory Concentration 50
High Pressure Liquid Chromatography

ASJC Scopus subject areas

  • Pharmacology

Cite this

Identification and characterization of human cytchrome P450 (CYP) isoforms interacting with cisapride. / Desta, Zeruesenay; Thacker, David; Soukhova, Nadia; Shin, Jae Gook; Flockhart, David A.

In: Clinical Pharmacology and Therapeutics, Vol. 65, No. 2, 1999, p. 126.

Research output: Contribution to journalArticle

Desta, Zeruesenay ; Thacker, David ; Soukhova, Nadia ; Shin, Jae Gook ; Flockhart, David A. / Identification and characterization of human cytchrome P450 (CYP) isoforms interacting with cisapride. In: Clinical Pharmacology and Therapeutics. 1999 ; Vol. 65, No. 2. pp. 126.
@article{649fd63d2b614ef997b590be7bc9aa37,
title = "Identification and characterization of human cytchrome P450 (CYP) isoforms interacting with cisapride",
abstract = "Using human liver microsomes (HLMs) and recombinant CYP450s, we characterized the metabolism of cisapride and documented its inhibitory effect on CYP450s. Two cisapride enantiomers (EI and EII) were separated and fractions collected for study. HPLC chromatograms of HLM incubates of cisapride separated 3 metabolite peaks, M1-M3. The identity of M1 is consistent with norcisapride, while M2 and M3 may represent oxidative hydroxylation of the fluorophenyl ring. The formation rate of M1 and 10μM cisapride in 15 HLMs significantly correlated with the activities of CYP3A, 2C19 and 1A2. Of CYP isoform specific inhibitors tested, ketoconazole potently inhibited the formation of M1, M2 and M3. Of the 10 recombinant CYP450s tested, CYP3A4, 2C8, 2C19, 2C9, 1A2 and 2B6 formed M1. Inhibition by racemic cisapride of CYP2D6 was relatively potent (Ki=14±16μM), of CYP1A2 (Ki=40±12μM) moderate and of CYP3A (Ki=142±27μM) weak. The inhibition by cisapride of CYP2D6 appears to be due to EII as the IC50 (μM) of EII (20±6) was closer to that of racemic cisapride (10±1.8), while that of EI (59±17) was 6-fold higher. In conclusion, cisapride is metabolized by multiple CYP450s in vitro which may make prediction of cisapride drug interaction and cardiac toxicity more difficult than described in the manufacturer's labels. Cisapride appears to inhibit CYP2D6-mediated metabolism in a stereoselective manner.",
author = "Zeruesenay Desta and David Thacker and Nadia Soukhova and Shin, {Jae Gook} and Flockhart, {David A.}",
year = "1999",
language = "English (US)",
volume = "65",
pages = "126",
journal = "Clinical Pharmacology and Therapeutics",
issn = "0009-9236",
publisher = "Nature Publishing Group",
number = "2",

}

TY - JOUR

T1 - Identification and characterization of human cytchrome P450 (CYP) isoforms interacting with cisapride

AU - Desta, Zeruesenay

AU - Thacker, David

AU - Soukhova, Nadia

AU - Shin, Jae Gook

AU - Flockhart, David A.

PY - 1999

Y1 - 1999

N2 - Using human liver microsomes (HLMs) and recombinant CYP450s, we characterized the metabolism of cisapride and documented its inhibitory effect on CYP450s. Two cisapride enantiomers (EI and EII) were separated and fractions collected for study. HPLC chromatograms of HLM incubates of cisapride separated 3 metabolite peaks, M1-M3. The identity of M1 is consistent with norcisapride, while M2 and M3 may represent oxidative hydroxylation of the fluorophenyl ring. The formation rate of M1 and 10μM cisapride in 15 HLMs significantly correlated with the activities of CYP3A, 2C19 and 1A2. Of CYP isoform specific inhibitors tested, ketoconazole potently inhibited the formation of M1, M2 and M3. Of the 10 recombinant CYP450s tested, CYP3A4, 2C8, 2C19, 2C9, 1A2 and 2B6 formed M1. Inhibition by racemic cisapride of CYP2D6 was relatively potent (Ki=14±16μM), of CYP1A2 (Ki=40±12μM) moderate and of CYP3A (Ki=142±27μM) weak. The inhibition by cisapride of CYP2D6 appears to be due to EII as the IC50 (μM) of EII (20±6) was closer to that of racemic cisapride (10±1.8), while that of EI (59±17) was 6-fold higher. In conclusion, cisapride is metabolized by multiple CYP450s in vitro which may make prediction of cisapride drug interaction and cardiac toxicity more difficult than described in the manufacturer's labels. Cisapride appears to inhibit CYP2D6-mediated metabolism in a stereoselective manner.

AB - Using human liver microsomes (HLMs) and recombinant CYP450s, we characterized the metabolism of cisapride and documented its inhibitory effect on CYP450s. Two cisapride enantiomers (EI and EII) were separated and fractions collected for study. HPLC chromatograms of HLM incubates of cisapride separated 3 metabolite peaks, M1-M3. The identity of M1 is consistent with norcisapride, while M2 and M3 may represent oxidative hydroxylation of the fluorophenyl ring. The formation rate of M1 and 10μM cisapride in 15 HLMs significantly correlated with the activities of CYP3A, 2C19 and 1A2. Of CYP isoform specific inhibitors tested, ketoconazole potently inhibited the formation of M1, M2 and M3. Of the 10 recombinant CYP450s tested, CYP3A4, 2C8, 2C19, 2C9, 1A2 and 2B6 formed M1. Inhibition by racemic cisapride of CYP2D6 was relatively potent (Ki=14±16μM), of CYP1A2 (Ki=40±12μM) moderate and of CYP3A (Ki=142±27μM) weak. The inhibition by cisapride of CYP2D6 appears to be due to EII as the IC50 (μM) of EII (20±6) was closer to that of racemic cisapride (10±1.8), while that of EI (59±17) was 6-fold higher. In conclusion, cisapride is metabolized by multiple CYP450s in vitro which may make prediction of cisapride drug interaction and cardiac toxicity more difficult than described in the manufacturer's labels. Cisapride appears to inhibit CYP2D6-mediated metabolism in a stereoselective manner.

UR - http://www.scopus.com/inward/record.url?scp=0000210089&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000210089&partnerID=8YFLogxK

M3 - Article

VL - 65

SP - 126

JO - Clinical Pharmacology and Therapeutics

JF - Clinical Pharmacology and Therapeutics

SN - 0009-9236

IS - 2

ER -