Effects of Genetic Variants on Carboxylesterase 1 Gene Expression, and Clopidogrel Pharmacokinetics and Antiplatelet Effects

Abstract

Several single nucleotide variations (SNVs) affect carboxylesterase 1 (CES1) activity, but the effects of genetic variants on CES1 gene expression have not been systematically investigated. Therefore, our aim was to investigate effects of genetic variants on CES1 gene expression in two independent whole blood sample cohorts of 192 (discovery) and 88 (replication) healthy volunteers and in a liver sample cohort of 177 patients. Furthermore, we investigated possible effects of the found variants on clopidogrel pharmacokinetics (n = 106) and pharmacodynamics (n = 46) in healthy volunteers, who had ingested a single 300 mg or 600 mg dose of clopidogrel. Using massively parallel sequencing, we discovered two CES1 SNVs, rs12443580 and rs8192935, to be strongly and independently associated with a 39% (p = 4.0 × 10 ) and 31% (p = 2.5 × 10 ) reduction in CES1 whole blood expression per copy of the minor allele. These findings were replicated in the replication cohort. However, these SNVs did not affect CES1 liver expression, or clopidogrel pharmacokinetics or pharmacodynamics. Conversely, the CES1 c.428G>A missense SNV (rs71647871) impaired the hydrolysis of clopidogrel, increased exposure to clopidogrel active metabolite and enhanced its antiplatelet effects. In conclusion, the rs12443580 and rs8192935 variants reduce CES1 expression in whole blood but not in the liver. These tissue-specific effects may result in substrate-dependent effects of the two SNVs on CES1-mediated drug metabolism.

Influence of Transporter Polymorphisms on Drug Disposition and Response: A Perspective From the International Transporter Consortium

“…The following are examples of how NGS have been used in the discovery of variants in membrane transporters, which are associated with drug disposition, response or toxicity.

NGS to identify genetic determinants for drug levels (or exposure). Recently, the Niemi laboratory used NGS to sequence candidate genes to identify the genetic basis of interindividual variation in response to montelukast and clopidogrel as well as concentrations of drug metabolites . These studies were conducted in ∼190 healthy volunteers, focusing on relevant enzymes and membrane transport proteins.

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“…NGS to identify genetic determinants for drug levels (or exposure). Recently, the Niemi laboratory used NGS to sequence candidate genes to identify the genetic basis of inter-individual variation in response to montelukast and clopidogrel as well as concentrations of drug metabolites 101,102 . These studies were conducted in approximately 190 healthy volunteers, focusing on relevant enzymes and membrane transport proteins.…”

Selective Carboxylesterase Inhibitors for Improving Efficacy, Safety and Rational use of Ester-Containing Drugs

“…В последние годы наблюдается лавинообразный рост числа публикаций по гидролитическому метаболизму с участием карбоксилэстераз (КЭ, КФ 3.1.1.1) -разнообразных по химической структуре лекарственных препаратов, содержащих сложноэфирную, карбаматную или амидную группировку. Это препараты различных фармакологических групп, отдельные представители которых показаны на рисунке 1 [1,2].…”

Pharmacokinetics and Bioequivalence of Clopidogrel Hydrogen Sulfate Tablets in Fed and Fasted Conditions: An Open‐Label, Randomized, Semireplicated Crossover Study in Healthy Chinese Volunteers

“…Clopidogrel is a prodrug that undergoes hepatic biotransformation to an active metabolite, which binds irreversibly to the P2Y12 receptor and inhibits ADP‐mediated platelet activation and aggregation. Around 85% of clopidogrel is hydrolyzed to the inactive metabolite clopidogrel carboxylic acid by esterases including carboxylesterase 1 5 . Clopidogrel carboxylic acid is metabolized mainly by UDP‐glucuronosyltransferase (UGT)2B7 and UGT2B4 in the liver and by UGT2B17 in the small intestinal wall 6 .…”

“…Around 85% of clopidogrel is hydrolyzed to the inactive metabolite clopidogrel carboxylic acid by esterases including carboxylesterase 1. 5 Clopidogrel carboxylic acid is metabolized mainly by UDPglucuronosyltransferase (UGT)2B7 and UGT2B4 in the liver and by UGT2B17 in the small intestinal wall. 6 Only 15% of clopidogrel is available for transformation into active metabolite through 2 sequential oxidative reactions by the cytochrome P450 system: the first step involves CYP1A2, CYP2B6, and CYP2C19, and the second step involves CYP2B6, CYP2C9, CYP2C19, CYP3A4, and CYP3A5.…”