Inhibition of human intestinal wall metabolism by macrolide antibiotics: Effect of clarithromycin on cytochrome P450 3A4/5 activity and expression

Amar G. Pinto, Ying Hong Wang, Naga Chalasani, Todd Skaar, Dhanashri Kolwankar, J. Christopher Gorski, Suthat Liangpunsakul, Mitchell A. Hamman, Million Arefayene, Stephen D. Hall

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Background: Clarithromycin increases both hepatic and intestinal availability of the selective cytochrome P450 (CYP) 3A probe midazolam. This study was designed to identify determinants of variability in the extent of intestinal wall CYP3A inhibition by clarithromycin, such as CYP3A5 genotype, and the mechanism of inhibition. Methods: Ten healthy volunteers received 500 mg oral clarithromycin twice a day for 7 days. Before and after administration of clarithromycin, small-bowel mucosal biopsy specimens were obtained endoscopically. Intestinal CYP3A activity was determined from the rate of 1′-hydroxymidazolam and 4-hydroxymidazolam formation by incubation of small-bowel homogenate with midazolam (25 μmol/L) and NADPH for 5 minutes. Intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid was quantified by real-time reverse transcriptase-polymerase chain reaction. Intestinal CYP3A4 and CYP3A5 protein concentrations were determined by immunoblotting. Serum and homogenate concentrations of midazolam, clarithromycin, and metabolites were determined by liquid chromatography-mass spectrometry. CYP3A5 genotype was determined by real-time polymerase chain reaction. Results: The formation of 1′-hydroxymidazolam (1.36 ± 0.46 pmol·min -1·mg-1 at baseline versus 0.35 ± 0.16 pmol·min-1·mg-1 after administration) and 4-hydroxymidazolam (0.39 ± 0.12 pmol·min -1·mg-1 at baseline versus 0.12 ± 0.05 pmol·min-1·mg-1 after administration) was significantly (P < .001) reduced after clarithromycin administration. Clarithromycin administration did not result in a significant change in intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid and protein expression. All subjects had detectable serum clarithromycin concentrations after 7 days of clarithromycin (3.71 ± 2.43 μmol/L). The mean concentration of clarithromycin in the intestinal biopsy homogenate was 1.2 ± 0.7 nmol/L (range, 0.42-2.39 nmol/L). Compared with CYP3A5 nonexpressers, subjects with at least 1 CYP3A5*1 allele (CYP3A5 expressers) had greater inhibition of intestinal CYP3A activity after treatment with clarithromycin. There was a strong linear relationship between the decrease in intestinal CYP3A activity and baseline catalytic activity (R2 = 0.9). Conclusion: Baseline intestinal activity of CYP3A4 was a key determinant of variability of the inhibitory effect of clarithromycin among individuals. CYP3A5*1 alleles were associated with greater baseline intestinal CYP3A activity and, therefore, greater extent of inhibition. The primary in vivo mechanism was not rapidly reversible competitive or irreversible inhibition but was likely formation of metabolic intermediate complexes.

Original languageEnglish
Pages (from-to)178-188
Number of pages11
JournalClinical Pharmacology and Therapeutics
Volume77
Issue number3
DOIs
StatePublished - Mar 2005

Fingerprint

Cytochrome P-450 CYP3A
Clarithromycin
Macrolides
Anti-Bacterial Agents
Midazolam
Real-Time Polymerase Chain Reaction
Alleles
Genotype
RNA
Biopsy

ASJC Scopus subject areas

  • Pharmacology

Cite this

Inhibition of human intestinal wall metabolism by macrolide antibiotics : Effect of clarithromycin on cytochrome P450 3A4/5 activity and expression. / Pinto, Amar G.; Wang, Ying Hong; Chalasani, Naga; Skaar, Todd; Kolwankar, Dhanashri; Gorski, J. Christopher; Liangpunsakul, Suthat; Hamman, Mitchell A.; Arefayene, Million; Hall, Stephen D.

In: Clinical Pharmacology and Therapeutics, Vol. 77, No. 3, 03.2005, p. 178-188.

Research output: Contribution to journalArticle

Pinto, Amar G. ; Wang, Ying Hong ; Chalasani, Naga ; Skaar, Todd ; Kolwankar, Dhanashri ; Gorski, J. Christopher ; Liangpunsakul, Suthat ; Hamman, Mitchell A. ; Arefayene, Million ; Hall, Stephen D. / Inhibition of human intestinal wall metabolism by macrolide antibiotics : Effect of clarithromycin on cytochrome P450 3A4/5 activity and expression. In: Clinical Pharmacology and Therapeutics. 2005 ; Vol. 77, No. 3. pp. 178-188.
@article{507dd37c65284c92ae581d317259919d,
title = "Inhibition of human intestinal wall metabolism by macrolide antibiotics: Effect of clarithromycin on cytochrome P450 3A4/5 activity and expression",
abstract = "Background: Clarithromycin increases both hepatic and intestinal availability of the selective cytochrome P450 (CYP) 3A probe midazolam. This study was designed to identify determinants of variability in the extent of intestinal wall CYP3A inhibition by clarithromycin, such as CYP3A5 genotype, and the mechanism of inhibition. Methods: Ten healthy volunteers received 500 mg oral clarithromycin twice a day for 7 days. Before and after administration of clarithromycin, small-bowel mucosal biopsy specimens were obtained endoscopically. Intestinal CYP3A activity was determined from the rate of 1′-hydroxymidazolam and 4-hydroxymidazolam formation by incubation of small-bowel homogenate with midazolam (25 μmol/L) and NADPH for 5 minutes. Intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid was quantified by real-time reverse transcriptase-polymerase chain reaction. Intestinal CYP3A4 and CYP3A5 protein concentrations were determined by immunoblotting. Serum and homogenate concentrations of midazolam, clarithromycin, and metabolites were determined by liquid chromatography-mass spectrometry. CYP3A5 genotype was determined by real-time polymerase chain reaction. Results: The formation of 1′-hydroxymidazolam (1.36 ± 0.46 pmol·min -1·mg-1 at baseline versus 0.35 ± 0.16 pmol·min-1·mg-1 after administration) and 4-hydroxymidazolam (0.39 ± 0.12 pmol·min -1·mg-1 at baseline versus 0.12 ± 0.05 pmol·min-1·mg-1 after administration) was significantly (P < .001) reduced after clarithromycin administration. Clarithromycin administration did not result in a significant change in intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid and protein expression. All subjects had detectable serum clarithromycin concentrations after 7 days of clarithromycin (3.71 ± 2.43 μmol/L). The mean concentration of clarithromycin in the intestinal biopsy homogenate was 1.2 ± 0.7 nmol/L (range, 0.42-2.39 nmol/L). Compared with CYP3A5 nonexpressers, subjects with at least 1 CYP3A5*1 allele (CYP3A5 expressers) had greater inhibition of intestinal CYP3A activity after treatment with clarithromycin. There was a strong linear relationship between the decrease in intestinal CYP3A activity and baseline catalytic activity (R2 = 0.9). Conclusion: Baseline intestinal activity of CYP3A4 was a key determinant of variability of the inhibitory effect of clarithromycin among individuals. CYP3A5*1 alleles were associated with greater baseline intestinal CYP3A activity and, therefore, greater extent of inhibition. The primary in vivo mechanism was not rapidly reversible competitive or irreversible inhibition but was likely formation of metabolic intermediate complexes.",
author = "Pinto, {Amar G.} and Wang, {Ying Hong} and Naga Chalasani and Todd Skaar and Dhanashri Kolwankar and Gorski, {J. Christopher} and Suthat Liangpunsakul and Hamman, {Mitchell A.} and Million Arefayene and Hall, {Stephen D.}",
year = "2005",
month = "3",
doi = "10.1016/j.clpt.2004.10.002",
language = "English",
volume = "77",
pages = "178--188",
journal = "Clinical Pharmacology and Therapeutics",
issn = "0009-9236",
publisher = "Nature Publishing Group",
number = "3",

}

TY - JOUR

T1 - Inhibition of human intestinal wall metabolism by macrolide antibiotics

T2 - Effect of clarithromycin on cytochrome P450 3A4/5 activity and expression

AU - Pinto, Amar G.

AU - Wang, Ying Hong

AU - Chalasani, Naga

AU - Skaar, Todd

AU - Kolwankar, Dhanashri

AU - Gorski, J. Christopher

AU - Liangpunsakul, Suthat

AU - Hamman, Mitchell A.

AU - Arefayene, Million

AU - Hall, Stephen D.

PY - 2005/3

Y1 - 2005/3

N2 - Background: Clarithromycin increases both hepatic and intestinal availability of the selective cytochrome P450 (CYP) 3A probe midazolam. This study was designed to identify determinants of variability in the extent of intestinal wall CYP3A inhibition by clarithromycin, such as CYP3A5 genotype, and the mechanism of inhibition. Methods: Ten healthy volunteers received 500 mg oral clarithromycin twice a day for 7 days. Before and after administration of clarithromycin, small-bowel mucosal biopsy specimens were obtained endoscopically. Intestinal CYP3A activity was determined from the rate of 1′-hydroxymidazolam and 4-hydroxymidazolam formation by incubation of small-bowel homogenate with midazolam (25 μmol/L) and NADPH for 5 minutes. Intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid was quantified by real-time reverse transcriptase-polymerase chain reaction. Intestinal CYP3A4 and CYP3A5 protein concentrations were determined by immunoblotting. Serum and homogenate concentrations of midazolam, clarithromycin, and metabolites were determined by liquid chromatography-mass spectrometry. CYP3A5 genotype was determined by real-time polymerase chain reaction. Results: The formation of 1′-hydroxymidazolam (1.36 ± 0.46 pmol·min -1·mg-1 at baseline versus 0.35 ± 0.16 pmol·min-1·mg-1 after administration) and 4-hydroxymidazolam (0.39 ± 0.12 pmol·min -1·mg-1 at baseline versus 0.12 ± 0.05 pmol·min-1·mg-1 after administration) was significantly (P < .001) reduced after clarithromycin administration. Clarithromycin administration did not result in a significant change in intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid and protein expression. All subjects had detectable serum clarithromycin concentrations after 7 days of clarithromycin (3.71 ± 2.43 μmol/L). The mean concentration of clarithromycin in the intestinal biopsy homogenate was 1.2 ± 0.7 nmol/L (range, 0.42-2.39 nmol/L). Compared with CYP3A5 nonexpressers, subjects with at least 1 CYP3A5*1 allele (CYP3A5 expressers) had greater inhibition of intestinal CYP3A activity after treatment with clarithromycin. There was a strong linear relationship between the decrease in intestinal CYP3A activity and baseline catalytic activity (R2 = 0.9). Conclusion: Baseline intestinal activity of CYP3A4 was a key determinant of variability of the inhibitory effect of clarithromycin among individuals. CYP3A5*1 alleles were associated with greater baseline intestinal CYP3A activity and, therefore, greater extent of inhibition. The primary in vivo mechanism was not rapidly reversible competitive or irreversible inhibition but was likely formation of metabolic intermediate complexes.

AB - Background: Clarithromycin increases both hepatic and intestinal availability of the selective cytochrome P450 (CYP) 3A probe midazolam. This study was designed to identify determinants of variability in the extent of intestinal wall CYP3A inhibition by clarithromycin, such as CYP3A5 genotype, and the mechanism of inhibition. Methods: Ten healthy volunteers received 500 mg oral clarithromycin twice a day for 7 days. Before and after administration of clarithromycin, small-bowel mucosal biopsy specimens were obtained endoscopically. Intestinal CYP3A activity was determined from the rate of 1′-hydroxymidazolam and 4-hydroxymidazolam formation by incubation of small-bowel homogenate with midazolam (25 μmol/L) and NADPH for 5 minutes. Intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid was quantified by real-time reverse transcriptase-polymerase chain reaction. Intestinal CYP3A4 and CYP3A5 protein concentrations were determined by immunoblotting. Serum and homogenate concentrations of midazolam, clarithromycin, and metabolites were determined by liquid chromatography-mass spectrometry. CYP3A5 genotype was determined by real-time polymerase chain reaction. Results: The formation of 1′-hydroxymidazolam (1.36 ± 0.46 pmol·min -1·mg-1 at baseline versus 0.35 ± 0.16 pmol·min-1·mg-1 after administration) and 4-hydroxymidazolam (0.39 ± 0.12 pmol·min -1·mg-1 at baseline versus 0.12 ± 0.05 pmol·min-1·mg-1 after administration) was significantly (P < .001) reduced after clarithromycin administration. Clarithromycin administration did not result in a significant change in intestinal CYP3A4 and CYP3A5 messenger ribonucleic acid and protein expression. All subjects had detectable serum clarithromycin concentrations after 7 days of clarithromycin (3.71 ± 2.43 μmol/L). The mean concentration of clarithromycin in the intestinal biopsy homogenate was 1.2 ± 0.7 nmol/L (range, 0.42-2.39 nmol/L). Compared with CYP3A5 nonexpressers, subjects with at least 1 CYP3A5*1 allele (CYP3A5 expressers) had greater inhibition of intestinal CYP3A activity after treatment with clarithromycin. There was a strong linear relationship between the decrease in intestinal CYP3A activity and baseline catalytic activity (R2 = 0.9). Conclusion: Baseline intestinal activity of CYP3A4 was a key determinant of variability of the inhibitory effect of clarithromycin among individuals. CYP3A5*1 alleles were associated with greater baseline intestinal CYP3A activity and, therefore, greater extent of inhibition. The primary in vivo mechanism was not rapidly reversible competitive or irreversible inhibition but was likely formation of metabolic intermediate complexes.

UR - http://www.scopus.com/inward/record.url?scp=13944250702&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=13944250702&partnerID=8YFLogxK

U2 - 10.1016/j.clpt.2004.10.002

DO - 10.1016/j.clpt.2004.10.002

M3 - Article

C2 - 15735612

AN - SCOPUS:13944250702

VL - 77

SP - 178

EP - 188

JO - Clinical Pharmacology and Therapeutics

JF - Clinical Pharmacology and Therapeutics

SN - 0009-9236

IS - 3

ER -