In vitro metabolism of montelukast by cytochrome P450S and UDP-glucuronosyltransferases

Josiane De Oliveira Cardoso, Regina Vincenzi Oliveira, Jessica Bo Li Lu, Zeruesenay Desta

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Montelukast has been recommended as a selective in vitro and in vivo probe of cytochrome P450 (P450) CYP2C8 activity, but its selectivity toward this enzyme remains unclear. We performed detailed characterization of montelukast metabolism in vitro using human liver microsomes (HLMs), expressed P450s, and uridine 59-diphospho-glucuronosyltransferases (UGTs). Kinetic and inhibition experiments performed at therapeutically relevant concentrations reveal that CYP2C8 and CYP2C9 are the principal enzymes responsible for montelukast 36-hydroxylation to 1,2-diol. CYP3A4 was the main catalyst of montelukast sulfoxidation and stereoselective 21-hydroxylation, and multiple P450s participated in montelukast 25-hydroxylation. We confirmed direct glucuronidation of montelukast to an acyl-glucuronide. We also identified a novel peak that appears consistent with an etherglucuronide. Kinetic analysis in HLMs and experiments in expressed UGTs indicate that both metabolites were exclusively formed by UGT1A3. Comparison of in vitro intrinsic clearance in HLMs suggest that direct glucuronidation may play a greater role in the overall metabolism of montelukast than does P450-mediated oxidation, but the in vivo contribution of UGT1A3 needs further testing. In conclusion, our in vitro findings provide new insight toward montelukast metabolism. The utility of montelukast as a probe of CYP2C8 activity may be compromised owing to involvement of multiple P450s and UGT1A3 in its metabolism.

Original languageEnglish (US)
Pages (from-to)1905-1916
Number of pages12
JournalDrug Metabolism and Disposition
Volume43
Issue number12
DOIs
StatePublished - Dec 1 2015

Fingerprint

montelukast
Glucuronosyltransferase
Cytochromes
Liver Microsomes
Hydroxylation
Uridine
In Vitro Techniques
Cytochrome P-450 CYP3A
Glucuronides

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Cite this

In vitro metabolism of montelukast by cytochrome P450S and UDP-glucuronosyltransferases. / De Oliveira Cardoso, Josiane; Oliveira, Regina Vincenzi; Lu, Jessica Bo Li; Desta, Zeruesenay.

In: Drug Metabolism and Disposition, Vol. 43, No. 12, 01.12.2015, p. 1905-1916.

Research output: Contribution to journalArticle

De Oliveira Cardoso, Josiane ; Oliveira, Regina Vincenzi ; Lu, Jessica Bo Li ; Desta, Zeruesenay. / In vitro metabolism of montelukast by cytochrome P450S and UDP-glucuronosyltransferases. In: Drug Metabolism and Disposition. 2015 ; Vol. 43, No. 12. pp. 1905-1916.
@article{a91445e5b8ae45d388db4b3a17fdd0f5,
title = "In vitro metabolism of montelukast by cytochrome P450S and UDP-glucuronosyltransferases",
abstract = "Montelukast has been recommended as a selective in vitro and in vivo probe of cytochrome P450 (P450) CYP2C8 activity, but its selectivity toward this enzyme remains unclear. We performed detailed characterization of montelukast metabolism in vitro using human liver microsomes (HLMs), expressed P450s, and uridine 59-diphospho-glucuronosyltransferases (UGTs). Kinetic and inhibition experiments performed at therapeutically relevant concentrations reveal that CYP2C8 and CYP2C9 are the principal enzymes responsible for montelukast 36-hydroxylation to 1,2-diol. CYP3A4 was the main catalyst of montelukast sulfoxidation and stereoselective 21-hydroxylation, and multiple P450s participated in montelukast 25-hydroxylation. We confirmed direct glucuronidation of montelukast to an acyl-glucuronide. We also identified a novel peak that appears consistent with an etherglucuronide. Kinetic analysis in HLMs and experiments in expressed UGTs indicate that both metabolites were exclusively formed by UGT1A3. Comparison of in vitro intrinsic clearance in HLMs suggest that direct glucuronidation may play a greater role in the overall metabolism of montelukast than does P450-mediated oxidation, but the in vivo contribution of UGT1A3 needs further testing. In conclusion, our in vitro findings provide new insight toward montelukast metabolism. The utility of montelukast as a probe of CYP2C8 activity may be compromised owing to involvement of multiple P450s and UGT1A3 in its metabolism.",
author = "{De Oliveira Cardoso}, Josiane and Oliveira, {Regina Vincenzi} and Lu, {Jessica Bo Li} and Zeruesenay Desta",
year = "2015",
month = "12",
day = "1",
doi = "10.1124/dmd.115.065763",
language = "English (US)",
volume = "43",
pages = "1905--1916",
journal = "Drug Metabolism and Disposition",
issn = "0090-9556",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "12",

}

TY - JOUR

T1 - In vitro metabolism of montelukast by cytochrome P450S and UDP-glucuronosyltransferases

AU - De Oliveira Cardoso, Josiane

AU - Oliveira, Regina Vincenzi

AU - Lu, Jessica Bo Li

AU - Desta, Zeruesenay

PY - 2015/12/1

Y1 - 2015/12/1

N2 - Montelukast has been recommended as a selective in vitro and in vivo probe of cytochrome P450 (P450) CYP2C8 activity, but its selectivity toward this enzyme remains unclear. We performed detailed characterization of montelukast metabolism in vitro using human liver microsomes (HLMs), expressed P450s, and uridine 59-diphospho-glucuronosyltransferases (UGTs). Kinetic and inhibition experiments performed at therapeutically relevant concentrations reveal that CYP2C8 and CYP2C9 are the principal enzymes responsible for montelukast 36-hydroxylation to 1,2-diol. CYP3A4 was the main catalyst of montelukast sulfoxidation and stereoselective 21-hydroxylation, and multiple P450s participated in montelukast 25-hydroxylation. We confirmed direct glucuronidation of montelukast to an acyl-glucuronide. We also identified a novel peak that appears consistent with an etherglucuronide. Kinetic analysis in HLMs and experiments in expressed UGTs indicate that both metabolites were exclusively formed by UGT1A3. Comparison of in vitro intrinsic clearance in HLMs suggest that direct glucuronidation may play a greater role in the overall metabolism of montelukast than does P450-mediated oxidation, but the in vivo contribution of UGT1A3 needs further testing. In conclusion, our in vitro findings provide new insight toward montelukast metabolism. The utility of montelukast as a probe of CYP2C8 activity may be compromised owing to involvement of multiple P450s and UGT1A3 in its metabolism.

AB - Montelukast has been recommended as a selective in vitro and in vivo probe of cytochrome P450 (P450) CYP2C8 activity, but its selectivity toward this enzyme remains unclear. We performed detailed characterization of montelukast metabolism in vitro using human liver microsomes (HLMs), expressed P450s, and uridine 59-diphospho-glucuronosyltransferases (UGTs). Kinetic and inhibition experiments performed at therapeutically relevant concentrations reveal that CYP2C8 and CYP2C9 are the principal enzymes responsible for montelukast 36-hydroxylation to 1,2-diol. CYP3A4 was the main catalyst of montelukast sulfoxidation and stereoselective 21-hydroxylation, and multiple P450s participated in montelukast 25-hydroxylation. We confirmed direct glucuronidation of montelukast to an acyl-glucuronide. We also identified a novel peak that appears consistent with an etherglucuronide. Kinetic analysis in HLMs and experiments in expressed UGTs indicate that both metabolites were exclusively formed by UGT1A3. Comparison of in vitro intrinsic clearance in HLMs suggest that direct glucuronidation may play a greater role in the overall metabolism of montelukast than does P450-mediated oxidation, but the in vivo contribution of UGT1A3 needs further testing. In conclusion, our in vitro findings provide new insight toward montelukast metabolism. The utility of montelukast as a probe of CYP2C8 activity may be compromised owing to involvement of multiple P450s and UGT1A3 in its metabolism.

UR - http://www.scopus.com/inward/record.url?scp=84948957404&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84948957404&partnerID=8YFLogxK

U2 - 10.1124/dmd.115.065763

DO - 10.1124/dmd.115.065763

M3 - Article

VL - 43

SP - 1905

EP - 1916

JO - Drug Metabolism and Disposition

JF - Drug Metabolism and Disposition

SN - 0090-9556

IS - 12

ER -