Rasmussen HB

References (25)

Title : Combined Ensemble Docking and Machine Learning in Identification of Therapeutic Agents with Potential Inhibitory Effect on Human CES1 - Briand_2019_Molecules_24_
Author(s) : Briand E , Thomsen R , Linnet K , Rasmussen HB , Brunak S , Taboureau O
Ref : Molecules , 24 : , 2019
Abstract : The human carboxylesterase 1 (CES1), responsible for the biotransformation of many diverse therapeutic agents, may contribute to the occurrence of adverse drug reactions and therapeutic failure through drug interactions. The present study is designed to address the issue of potential drug interactions resulting from the inhibition of CES1. Based on an ensemble of 10 crystal structures complexed with different ligands and a set of 294 known CES1 ligands, we used docking (Autodock Vina) and machine learning methodologies (LDA, QDA and multilayer perceptron), considering the different energy terms from the scoring function to assess the best combination to enable the identification of CES1 inhibitors. The protocol was then applied on a library of 1114 FDA-approved drugs and eight drugs were selected for in vitro CES1 inhibition. An inhibition effect was observed for diltiazem (IC50 = 13.9 microM). Three others drugs (benztropine, iloprost and treprostinil), exhibited a weak CES1 inhibitory effects with IC50 values of 298.2 microM, 366.8 microM and 391.6 microM respectively. In conclusion, the binding site of CES1 is relatively flexible and can adapt its conformation to different types of ligands. Combining ensemble docking and machine learning approaches improves the prediction of CES1 inhibitors compared to a docking study using only one crystal structure.
ESTHER : Briand_2019_Molecules_24_
PubMedSearch : Briand_2019_Molecules_24_
PubMedID: 31362390

Title : The impact of human CES1 genetic variation on enzyme activity assessed by ritalinic acid\/methylphenidate ratios - Stage_2019_Basic.Clin.Pharmacol.Toxicol_125_54
Author(s) : Stage C , Dalhoff K , Rasmussen HB , Schow Guski L , Thomsen R , Bjerre D , Ferrero-Miliani L , Busk Madsen M , Jurgens G
Ref : Basic Clin Pharmacol Toxicol , 125 :54 , 2019
Abstract : The present clinical trial investigated the impact of selected SNPs in CES1 on the metabolic activity of the enzyme. For this purpose, we used methylphenidate (MPH) as a pharmacological probe and the d-RA/d-MPH (metabolite/parent drug) ratios as a measure of enzymatic activity. This metabolic ratio (MR) was validated against the AUC ratios (AUCd -RA /AUCd -MPH ). CES1 SNPs from 120 volunteers were identified, and 12 SNPs fulfilling predefined inclusion criteria were analysed separately, comparing the effect of each genotype on the metabolic ratios. The SNP criteria were as follows: presence of Hardy-Weinberg equilibrium, a minor allele frequency >/= 0.01 and a clearly interpretable sequencing result in at least 30% of the individuals. Each participant ingested 10 mg of racemic methylphenidate, and blood samples were drawn prior to and 3 hours after drug administration. The SNP analysis confirmed the considerable impact of rs71647871 (G143E) in exon 4 on drug metabolism. In addition, three volunteers with markedly lower median MR, indicating decreased CES1 activity, harboured the same combination of three SNPs in intron 5. The median MR for these SNPs was 8.2 for the minor allele compared to 16.4 for the major alleles (P = 0.04). Hence, one of these or the combination of these SNPs could be of clinical significance considering that the median MR of the G143E group was 5.4. The precise genetic relationship of this finding is currently unknown, as is the clinical significance.
ESTHER : Stage_2019_Basic.Clin.Pharmacol.Toxicol_125_54
PubMedSearch : Stage_2019_Basic.Clin.Pharmacol.Toxicol_125_54
PubMedID: 30801959
Gene_locus related to this paper: human-CES1

Title : Carboxylesterase 1 genes: systematic review and evaluation of existing genotyping procedures - Rasmussen_2018_Drug.Metab.Pers.Ther_33_3
Author(s) : Rasmussen HB , Madsen MB
Ref : Drug Metab Pers Ther , 33 :3 , 2018
Abstract : The carboxylesterase 1 gene (CES1) encodes a hydrolase that metabolizes commonly used drugs. The CES1-related pseudogene, carboxylesterase 1 pseudogene 1 (CES1P1), has been implicated in gene exchange with CES1 and in the formation of hybrid genes including the carboxylesterase 1A2 gene (CES1A2). Hence, the CES1 region is complex. Using in silico PCR and alignment, we assessed the specificity of PCR-assisted procedures for genotyping CES1, CES1A2 and CES1P1 in studies identified in PubMed. We identified 33 such studies and excluded those that were not the first to use a procedure or lacked sequence information. After this 17 studies remained. Ten of these used haplotype-specific amplification, restriction enzyme treatment or amplicon sequencing, and included five that were predicted to lack specificity. All procedures for genotyping of single nucleotide polymorphisms in eight studies lacked specificity. One of these studies also used amplicon sequencing, thus being present in the group above. Some primers and their intended targets were mismatched. We provide experimental evidence that one of the procedures lacked specificity. Additionally, a complex pattern of segmental duplications in the CES1 region was revealed. In conclusion, many procedures for CES1, CES1A2 and CES1P1 genotyping appear to lack specificity. Knowledge about the segmental duplications may improve the typing of these genes.
ESTHER : Rasmussen_2018_Drug.Metab.Pers.Ther_33_3
PubMedSearch : Rasmussen_2018_Drug.Metab.Pers.Ther_33_3
PubMedID: 29427553
Gene_locus related to this paper: human-CES1

Title : Pharmacometabolomics Informs About Pharmacokinetic Profile of Methylphenidate - Kaddurah-Daouk_2018_CPT.Pharmacometrics.Syst.Pharmacol_7_525
Author(s) : Kaddurah-Daouk R , Hankemeier T , Scholl EH , Baillie R , Harms A , Stage C , Dalhoff KP , Jurgens G , Taboureau O , Nzabonimpa GS , Motsinger-Reif AA , Thomsen R , Linnet K , Rasmussen HB
Ref : CPT Pharmacometrics Syst Pharmacol , 7 :525 , 2018
Abstract : Carboxylesterase 1 (CES1) metabolizes methylphenidate and other drugs. CES1 gene variation only partially explains pharmacokinetic (PK) variability. Biomarkers predicting the PKs of drugs metabolized by CES1 are needed. We identified lipids in plasma from 44 healthy subjects that correlated with CES1 activity as determined by PK parameters of methylphenidate including a ceramide (q value = 0.001) and a phosphatidylcholine (q value = 0.005). Carriers of the CES1 143E allele had decreased methylphenidate metabolism and altered concentration of this phosphatidylcholine (q value = 0.040) and several high polyunsaturated fatty acid lipids (PUFAs). The half-maximal inhibitory concentration (IC50 ) values of chenodeoxycholate and taurocholate were 13.55 and 19.51 muM, respectively, consistent with a physiological significance. In silico analysis suggested that bile acid inhibition of CES1 involved both binding to the active and superficial sites of the enzyme. We initiated identification of metabolites predicting PKs of drugs metabolized by CES1 and suggest lipids to regulate or be regulated by this enzyme.
ESTHER : Kaddurah-Daouk_2018_CPT.Pharmacometrics.Syst.Pharmacol_7_525
PubMedSearch : Kaddurah-Daouk_2018_CPT.Pharmacometrics.Syst.Pharmacol_7_525
PubMedID: 30169917

Title : Nomenclature for alleles of the human carboxylesterase 1 gene -
Author(s) : Rasmussen HB , Madsen MB , Hansen PR
Ref : Pharmacogenet Genomics , 27 :78 , 2017
PubMedID: 27831961
Gene_locus related to this paper: human-CES1

Title : Carboxylesterase 1A2 encoding gene with increased transcription and potential rapid drug metabolism in Asian populations - Rasmussen_2017_Drug.Metab.Pers.Ther_32_163
Author(s) : Rasmussen HB , Madsen MB , Lyauk YK , Hansen PR , Hughes T
Ref : Drug Metab Pers Ther , 32 :163 , 2017
Abstract : The carboxylesterase 1 gene (CES1) encodes a hydrolase implicated in the metabolism of commonly used drugs. CES1A2, a hybrid of CES1 and a CES1-like pseudogene, has a promoter that is weak in most individuals. However, some individuals harbor a promoter haplotype of this gene with two overlapping Sp1 sites that confer significantly increased transcription potentially leading to rapid drug metabolism. This CES1A2 haplotype has previously been reported to be common among Asians. Using polymerase chain reaction followed by sequencing, the present study examined variation in the promoter and 5' untranslated region of CES1A2 in 120 Han Chinese and 120 Japanese people enrolled in the 1000 Genomes Project. We identified 11 single nucleotide variations, two of which were novel, in 145 of the individuals who were found to carry CES1A2. Alignment analysis indicated that the CES1A2 haplotype with the overlapping Sp1 sites has been generated by incorporation of a segment of CES1. All minor allele frequencies were equal to or below 0.022 and the frequencies of the minor haplotypes were up to 40-fold lower than previously reported, including that of the haplotype with the overlapping Sp1 sites. This information is novel and suggests that the pharmacogenetic relevance of CES1A2 is limited in Asians.
ESTHER : Rasmussen_2017_Drug.Metab.Pers.Ther_32_163
PubMedSearch : Rasmussen_2017_Drug.Metab.Pers.Ther_32_163
PubMedID: 28803225

Title : Reappraisal of the genetic diversity and pharmacogenetic assessment of CES1 - Ferrero-Miliani_2017_Pharmacogenomics_18_1241
Author(s) : Ferrero-Miliani L , Bjerre D , Stage C , Madsen MB , Jrgens G , Dalhoff KP , Rasmussen HB
Ref : Pharmacogenomics , 18 :1241 , 2017
Abstract : The CES1 gene encodes a hydrolase that metabolizes important drugs. Variants generated by exchange of segments with CES1P1 complicate genotyping of CES1. Using a highly specific procedure we examined DNA samples from 200 Caucasians and identified 46 single nucleotide variants (SNVs) in CES1 and 21 SNVs in CES1A2, a hybrid composed of CES1 and CES1P1. Several of these SNVs were novel. The frequencies of SNVs with a potential functional impact were below 0.02 suggesting limited pharmacogenetic potential for CES1 genotyping. In silico PCR revealed that the majority of the primer pairs for amplification of CES1 or CES1A2 in three previous studies lacked specificity, which partially explains a limited overlap with our findings.
ESTHER : Ferrero-Miliani_2017_Pharmacogenomics_18_1241
PubMedSearch : Ferrero-Miliani_2017_Pharmacogenomics_18_1241
PubMedID: 28786738

Title : The impact of CES1 genotypes on the pharmacokinetics of methylphenidate in healthy Danish subjects - Stage_2017_Br.J.Clin.Pharmacol_83_1506
Author(s) : Stage C , Jurgens G , Guski LS , Thomsen R , Bjerre D , Ferrero-Miliani L , Lyauk YK , Rasmussen HB , Dalhoff K
Ref : British Journal of Clinical Pharmacology , 83 :1506 , 2017
Abstract : AIMS: This study investigated the influence of CES1 variations, including the single nucleotide polymorphism (SNP) rs71647871 (G143E) and variation in copy number, on the pharmacokinetics of a single oral dose of 10 mg methylphenidate.
METHODS: CES1 genotype was obtained from 200 healthy Danish Caucasian volunteers. Based on the genotype, 44 (19 males and 25 females) were invited to participate in an open, prospective trial involving six predefined genotypes: three groups with two, three and four CES1 copies, respectively; a group of carriers of the CES1 143E allele; a group of individuals homozygous for CES1A1c (CES1VAR); and a group having three CES1 copies, in which the duplication, CES1A2, had increased transcriptional activity. Plasma concentrations of methylphenidate and its primary metabolites were determined at scheduled time points.
RESULTS: Median AUC of d-methylphenidate was significantly larger in the group carrying the 143E allele (53.3 ng ml-1 h-1 , range 38.6-93.9) than in the control group (21.4 ng ml-1 h-1 , range 15.7-34.9) (P < 0.0001). Median AUC of d-methylphenidate was significantly larger in the group with four CES1 copies (34.5 ng ml-1 h-1 , range 21.3-62.8) than in the control group (P = 0.01) and the group with three CES1 copies (23.8 ng ml-1 h-1 , range 15.3-32.0, P = 0.03). There was no difference between the groups with two and three copies of CES1.
CONCLUSIONS: The 143E allele resulted in an increased AUC, suggesting a significantly decreased CES1 enzyme activity. Surprisingly, this was also the case in subjects with homozygous duplication of CES1, perhaps reflecting an undiscovered mutation affecting the activity of the enzyme.
ESTHER : Stage_2017_Br.J.Clin.Pharmacol_83_1506
PubMedSearch : Stage_2017_Br.J.Clin.Pharmacol_83_1506
PubMedID: 28087982
Gene_locus related to this paper: human-CES1

Title : The Pharmacokinetics of Enalapril in Relation to CES1 Genotype in Healthy Danish Volunteers - Stage_2017_Basic.Clin.Pharmacol.Toxicol_121_487
Author(s) : Stage C , Jurgens G , Guski LS , Thomsen R , Bjerre D , Ferrero-Miliani L , Lyauk YK , Rasmussen HB , Dalhoff K
Ref : Basic Clin Pharmacol Toxicol , 121 :487 , 2017
Abstract : This study investigated the influence of variations in the carboxylesterase 1 gene (CES1) on the pharmacokinetics of enalapril. Forty-three healthy, Danish, Caucasian volunteers representing different pre-defined genotypes each received 10 mg of enalapril. At specified time-points, plasma concentrations of enalapril and the active metabolite enalaprilat were measured. The volunteers were divided into six different groups according to their genetic profile of CES1: group 1 (control group, n = 16) with two CES1 copies without non-synonymous SNPs in the exons; group 2 (n = 5) with four copies of CES1; group 3 (n = 6) harbouring the G143E polymorphism; group 4 (n = 2) with three CES1 copies and increased transcriptional activity of the duplication (CES1A2); group 5 (n = 4) harbouring the CES1A1c variant; and group 6 (n = 10) with three CES1 copies and the common promoter with low transcriptional activity of the duplication. The median AUC of enalaprilat in the control group was not significantly different from any of the other five groups (297 ng/ml x h in the control group versus 310, 282, 294, 344 and 306 ng/ml x h in groups 2-6, respectively). The terminal half-life of enalaprilat was significantly longer in group 6 compared with the control group (26.7 hr versus 12.7 hr, respectively). However, this was not considered clinically relevant. In conclusion, none of the selected variations of CES1 had a clinically relevant impact on the metabolism of enalapril.
ESTHER : Stage_2017_Basic.Clin.Pharmacol.Toxicol_121_487
PubMedSearch : Stage_2017_Basic.Clin.Pharmacol.Toxicol_121_487
PubMedID: 28639420
Gene_locus related to this paper: human-CES1

Title : Novel procedure with improved resolution and specificity for amplification and differentiation of variants of the gene encoding carboxylesterase 1 - Bjerre_2017_Pharmacogenet.Genomics_27_155
Author(s) : Bjerre D , Rasmussen HB
Ref : Pharmacogenet Genomics , 27 :155 , 2017
Abstract : Carboxylesterase 1 (CES1) is implicated in the metabolism of several commonly used drugs and other xenobiotics. The gene encoding this enzyme, CES1, is duplicated in some individuals. The original gene copy is called CES1A1. The duplicated version, CES1A2, is a hybrid of CES1A1 and the CES1-related pseudogene, CES1P1. Variants of CES1A2 with a weak and a strong promoter, respectively, have been reported. In addition, there are chimeric subtypes of CES1A1 that contain a segment of CES1P1. Collectively, this represents challenges to the genotyping of CES1 that previous procedures have had difficulties in solving, frequently leading to loss of specificity and inaccurate genotyping. Here, we report a novel and specific procedure that can selectively amplify CES1A1 and CES1A2 and accurately determine their variants. This procedure may be useful for personalization of treatments with drugs metabolized by CES1.
ESTHER : Bjerre_2017_Pharmacogenet.Genomics_27_155
PubMedSearch : Bjerre_2017_Pharmacogenet.Genomics_27_155
PubMedID: 28146010
Gene_locus related to this paper: human-CES1

Title : Population Pharmacokinetics of Methylphenidate in Healthy Adults Emphasizing Novel and Known Effects of Several Carboxylesterase 1 (CES1) Variants - Lyauk_2016_Clin.Transl.Sci_9_337
Author(s) : Lyauk YK , Stage C , Bergmann TK , Ferrero-Milliani L , Bjerre D , Thomsen R , Dalhoff KP , Rasmussen HB , Jurgens G
Ref : Clin Transl Sci , 9 :337 , 2016
Abstract : The aim of this study was to identify demographic and genetic factors that significantly affect methylphenidate (MPH) pharmacokinetics (PK), and may help explain interindividual variability and further increase the safety of MPH. d-MPH plasma concentrations, demographic covariates, and carboxylesterase 1 (CES1) genotypes were gathered from 122 healthy adults and analyzed using nonlinear mixed effects modeling. The structural model that best described the data was a two-compartment disposition model with absorption transit compartments. Novel effects of rs115629050 and CES1 diplotypes, as well as previously reported effects of rs71647871 and body weight, were included in the final model. Assessment of the independent and combined effect of CES1 covariates identified several specific risk factors that may result in severely increased d-MPH plasma exposure.
ESTHER : Lyauk_2016_Clin.Transl.Sci_9_337
PubMedSearch : Lyauk_2016_Clin.Transl.Sci_9_337
PubMedID: 27754602
Gene_locus related to this paper: human-CES1

Title : Pharmacodynamic Impact of Carboxylesterase 1 Gene Variants in Patients with Congestive Heart Failure Treated with Angiotensin-Converting Enzyme Inhibitors - Nelveg-Kristensen_2016_PLoS.One_11_e0163341
Author(s) : Nelveg-Kristensen KE , Bie P , Ferrero L , Bjerre D , Bruun NE , Egfjord M , Rasmussen HB , Hansen PR
Ref : PLoS ONE , 11 :e0163341 , 2016
Abstract : BACKGROUND: Variation in the carboxylesterase 1 gene (CES1) may contribute to the efficacy of ACEIs. Accordingly, we examined the impact of CES1 variants on plasma angiotensin II (ATII)/angiotensin I (ATI) ratio in patients with congestive heart failure (CHF) that underwent ACEI dose titrations. Five of these variants have previously been associated with drug response or increased CES1 expression, i.e., CES1 copy number variation, the variant of the duplicated CES1 gene with high transcriptional activity, rs71647871, rs2244613, and rs3815583. Additionally, nine variants, representatives of CES1Var, and three other CES1 variants were examined.
METHODS: Patients with CHF, and clinical indication for ACEIs were categorized according to their CES1 genotype. Differences in mean plasma ATII/ATI ratios between genotype groups after ACEI dose titration, expressed as the least square mean (LSM) with 95% confidence intervals (CIs), were assessed by analysis of variance.
RESULTS: A total of 200 patients were recruited and 127 patients (63.5%) completed the study. The mean duration of the CHF drug dose titration was 6.2 (SD 3.6) months. After ACEI dose titration, there was no difference in mean plasma ATII/ATI ratios between subjects with the investigated CES1 variants, and only one previously unexplored variation (rs2302722) qualified for further assessment. In the fully adjusted analysis of effects of rs2302722 on plasma ATII/ATI ratios, the difference in mean ATII/ATI ratio between the GG genotype and the minor allele carriers (GT and TT) was not significant, with a relative difference in LSMs of 0.67 (95% CI 0.43-1.07; P = 0.10). Results of analyses that only included enalapril-treated patients remained non-significant after Bonferroni correction for multiple parallel comparisons (difference in LSM 0.60 [95% CI 0.37-0.98], P = 0.045). CONCLUSION: These findings indicate that the included single variants of CES1 do not significantly influence plasma ATII/ATI ratios in CHF patients treated with ACEIs and are unlikely to be primary determinants of ACEI efficacy.
ESTHER : Nelveg-Kristensen_2016_PLoS.One_11_e0163341
PubMedSearch : Nelveg-Kristensen_2016_PLoS.One_11_e0163341
PubMedID: 27662362

Title : Investigating the impact of missense mutations in hCES1 by in silico structure-based approaches - Nzabonimpa_2016_Drug.Metab.Pers.Ther__
Author(s) : Nzabonimpa GS , Rasmussen HB , Brunak S , Taboureau O
Ref : Drug Metab Pers Ther , : , 2016
Abstract : Genetic variations in drug-metabolizing enzymes have been reported to influence pharmacokinetics, drug dosage, and other aspects that affect therapeutic outcomes. Most particularly, non-synonymous single-nucleotide polymorphisms (nsSNPs) resulting in amino acid changes disrupt potential functional sites responsible for protein activity, structure, or stability, which can account for individual susceptibility to disease and drug response. Investigating the impact of nsSNPs at a protein's structural level is a key step in understanding the relationship between genetic variants and the resulting phenotypic changes. For this purpose, in silico structure-based approaches have proven their relevance in providing an atomic-level description of the underlying mechanisms. The present review focuses on nsSNPs in human carboxylesterase 1 (hCES1), an enzyme involved in drug metabolism. We highlight how prioritization of functional nsSNPs through computational prediction techniques in combination with structure-based approaches, namely molecular docking and molecular dynamics simulations, is a powerful tool in providing insight into the underlying molecular mechanisms of nsSNPs phenotypic effects at microscopic level. Examples of in silico studies of carboxylesterases (CESs) are discussed, ranging from exploring the effect of mutations on enzyme activity to predicting the metabolism of new hCES1 substrates as well as to guiding rational design of CES-selective inhibitors.
ESTHER : Nzabonimpa_2016_Drug.Metab.Pers.Ther__
PubMedSearch : Nzabonimpa_2016_Drug.Metab.Pers.Ther__
PubMedID: 26900165
Gene_locus related to this paper: human-CES1

Title : Prognostic impact of carboxylesterase 1 gene variants in patients with congestive heart failure treated with angiotensin-converting enzyme inhibitors - Nelveg-Kristensen_2016_Pharmacogenet.Genomics_26_169
Author(s) : Nelveg-Kristensen KE , Madsen MB , Torp-Pedersen C , Kober L , Egfjord M , Hansen T , Pedersen O , Rasmussen HB , Hansen PR
Ref : Pharmacogenet Genomics , 26 :169 , 2016
Abstract : OBJECTIVE: Most angiotensin-converting enzyme inhibitors (ACEIs) are prodrugs activated by carboxylesterase 1 (CES1). We investigated the prognostic importance of CES1 gene (CES1) copy number variation and the rs3815583 single-nucleotide polymorphism in CES1 among ACEI-treated patients with congestive heart failure (CHF). METHODS: Danish patients with chronic CHF enrolled in the previously reported Echocardiography and Heart Outcome Study were categorized according to their CES1 variants and followed up for up to 10 years. Risk for cardiovascular death and all-cause death was modeled by Cox proportional hazard analyses. RESULTS: A total of 491 ACEI-treated patients were included in the analyses. After a mean follow-up of 5.5 years, we found no difference in the risk for cardiovascular death and all-cause death between patients having three [hazard ratios (HRs) 1.06 (95% confidence interval (CI) 0.77-1.45) and 1.16 (95% CI 0.88-1.52)] or four [HRs 0.88 (95% CI 0.39-2.01) and 1.37 (95% CI 0.74-2.54)] CES1 copies and those with two copies, respectively. Similarly, no difference in the risk for cardiovascular and all-cause death was found for patients heterozygous [HRs 0.91 (95% CI 0.70-1.19) and 0.88 (95% CI 0.69-1.12)] or homozygous [HRs 0.58 (95% CI 0.30-1.15) and 0.82 (95% CI 0.48-1.39)] for the rs3815583 minor allele versus patients homozygous for the major allele. The active promoter of CES1A2 and the rs71647871 single-nucleotide polymorphism minor allele were detected at very low frequencies. CONCLUSION: This study did not support the use of CES1 copy number variation or rs3815583 as a predictor of fatal outcomes in ACEI-treated patients with CHF.
ESTHER : Nelveg-Kristensen_2016_Pharmacogenet.Genomics_26_169
PubMedSearch : Nelveg-Kristensen_2016_Pharmacogenet.Genomics_26_169
PubMedID: 26761119
Gene_locus related to this paper: human-CES1

Title : Synthetic cannabimimetic agents metabolized by carboxylesterases - Thomsen_2015_Drug.Test.Anal_7_565
Author(s) : Thomsen R , Nielsen LM , Holm NB , Rasmussen HB , Linnet K
Ref : Drug Test Anal , 7 :565 , 2015
Abstract : Synthetic cannabimimetic agents are a large group of diverse compounds which act as agonists at cannabinoid receptors. Since 2004, synthetic cannabinoids have been used recreationally, although several of the compounds have been shown to cause severe toxicity in humans. In this study, the metabolism of two indazole carboxamide derivatives, AB-PINACA and AB-FUBINACA, was investigated by using human liver microsomes (HLM). For both compounds, a major metabolic pathway was the enzymatic hydrolysis of the primary amide, resulting in the major metabolites AB-PINACA-COOH and AB-FUBINACA-COOH. Other major metabolic pathways were mono-hydroxylation of the N-pentyl chain in AB-PINACA and mono-hydroxylation of the 1-amino-3-methyl-1-oxobutane moiety in AB-FUBINACA. To identify the enzyme(s) responsible for the amide hydrolysis, incubations with recombinant carboxylesterases and human serum, as well as inhibition studies in HLM and human pulmonary microsomes (HPM) were performed. Carboxylesterase 1 (CES1) was identified as the major human hepatic and pulmonary enzyme responsible for the amide hydrolysis.We employed similar studies to identify the esterase(s) involved in the previously described hydrolytic metabolism of two quinolineindole synthetic cannabinoids, PB-22 and 5F-PB-22, as well as the closely related compound, BB-22. Our investigations again revealed CES1 to be the key enzyme catalyzing these reactions. The identified major metabolites of AB-PINACA and AB-FUBINACA are likely to be useful in documenting drug usage in forensic and clinical screening. Additionally, the identification of CES1 as the main enzyme hydrolyzing these compounds improves our knowledge in the emerging field of xenobiotic metabolism by esterases. Copyright (c) 2014 John Wiley & Sons, Ltd.
ESTHER : Thomsen_2015_Drug.Test.Anal_7_565
PubMedSearch : Thomsen_2015_Drug.Test.Anal_7_565
PubMedID: 25346527

Title : Individualization of treatments with drugs metabolized by CES1: combining genetics and metabolomics - Rasmussen_2015_Pharmacogenomics_16_649
Author(s) : Rasmussen HB , Bjerre D , Linnet K , Jurgens G , Dalhoff K , Stefansson H , Hankemeier T , Kaddurah-Daouk R , Taboureau O , Brunak S , Houmann T , Jeppesen P , Pagsberg AK , Plessen K , Dyrborg J , Hansen PR , Hansen PE , Hughes T , Werge T
Ref : Pharmacogenomics , 16 :649 , 2015
Abstract : CES1 is involved in the hydrolysis of ester group-containing xenobiotic and endobiotic compounds including several essential and commonly used drugs. The individual variation in the efficacy and tolerability of many drugs metabolized by CES1 is considerable. Hence, there is a large interest in individualizing the treatment with these drugs. The present review addresses the issue of individualized treatment with drugs metabolized by CES1. It describes the composition of the gene encoding CES1, reports variants of this gene with focus upon those with a potential effect on drug metabolism and provides an overview of the protein structure of this enzyme bringing notice to mechanisms involved in the regulation of enzyme activity. Subsequently, the review highlights drugs metabolized by CES1 and argues that individual differences in the pharmacokinetics of these drugs play an important role in determining drug response and tolerability suggesting prospects for individualized drug therapies. Our review also discusses endogenous substrates of CES1 and assesses the potential of using metabolomic profiling of blood to identify proxies for the hepatic activity of CES1 that predict the rate of drug metabolism. Finally, the combination of genetics and metabolomics to obtain an accurate prediction of the individual response to CES1-dependent drugs is discussed.
ESTHER : Rasmussen_2015_Pharmacogenomics_16_649
PubMedSearch : Rasmussen_2015_Pharmacogenomics_16_649
PubMedID: 25896426

Title : Clopidogrel Bioactivation and Risk of Bleeding in Patients Cotreated With Angiotensin-Converting Enzyme Inhibitors After Myocardial Infarction: A Proof-of-Concept Study - Kristensen_2014_Clin.Pharmacol.Ther_96_713
Author(s) : Kristensen KE , Zhu HJ , Wang X , Gislason GH , Torp-Pedersen C , Rasmussen HB , Markowitz JS , Hansen PR
Ref : Clinical Pharmacology & Therapeutics , 96 :713 , 2014
Abstract : Clopidogrel is an oral antiplatelet prodrug, the majority of which is hydrolyzed to an inactive metabolite by hepatic carboxylesterase 1 (CES1). Most angiotensin-converting enzyme inhibitors (ACEIs) are also metabolized by this enzyme. We examined the effects of ACEIs on clopidogrel bioactivation in vitro and linked the results with a pharmacoepidemiological study. In vitro, ACEIs inhibited CES1-mediated hydrolysis of a model substrate, and trandolapril and enalapril increased formation of clopidogrel active metabolite. In 70,934 patients with myocardial infarction, hazard ratios for clinically significant bleeding in ACEI-treated patients cotreated with or without clopidogrel were 1.10 (95% confidence interval (CI): 0.97-1.25, P = 0.124) and 0.90 (95% CI: 0.81-0.99, P = 0.025), respectively, as compared with patients who did not receive ACEIs. This difference was statistically significant (P = 0.002). We conclude that cotreatment with selected ACEIs and clopidogrel may increase the risk of bleeding. Combination of in vitro and pharmacoepidemiological studies may be a useful paradigm for assessment of drug-drug interactions.
ESTHER : Kristensen_2014_Clin.Pharmacol.Ther_96_713
PubMedSearch : Kristensen_2014_Clin.Pharmacol.Ther_96_713
PubMedID: 25222620

Title : In vitro drug metabolism by human carboxylesterase 1: focus on Angiotensin-converting enzyme inhibitors - Thomsen_2014_Drug.Metab.Dispos_42_126
Author(s) : Thomsen R , Rasmussen HB , Linnet K
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 42 :126 , 2014
Abstract : Carboxylesterase 1 (CES1) is the major hydrolase in human liver. The enzyme is involved in the metabolism of several important therapeutic agents, drugs of abuse, and endogenous compounds. However, no studies have described the role of human CES1 in the activation of two commonly prescribed angiotensin-converting enzyme inhibitors: enalapril and ramipril. Here, we studied recombinant human CES1- and CES2-mediated hydrolytic activation of the prodrug esters enalapril and ramipril, compared with the activation of the known substrate trandolapril. Enalapril, ramipril, and trandolapril were readily hydrolyzed by CES1, but not by CES2. Ramipril and trandolapril exhibited Michaelis-Menten kinetics, while enalapril demonstrated substrate inhibition kinetics. Intrinsic clearances were 1.061, 0.360, and 0.02 ml/min/mg protein for ramipril, trandolapril, and enalapril, respectively. Additionally, we screened a panel of therapeutic drugs and drugs of abuse to assess their inhibition of the hydrolysis of p-nitrophenyl acetate by recombinant CES1 and human liver microsomes. The screening assay confirmed several known inhibitors of CES1 and identified two previously unreported inhibitors: the dihydropyridine calcium antagonist, isradipine, and the immunosuppressive agent, tacrolimus. CES1 plays a role in the metabolism of several drugs used in the treatment of common conditions, including hypertension, congestive heart failure, and diabetes mellitus; thus, there is a potential for clinically relevant drug-drug interactions. The findings in the present study may contribute to the prediction of such interactions in humans, thus opening up possibilities for safer drug treatments.
ESTHER : Thomsen_2014_Drug.Metab.Dispos_42_126
PubMedSearch : Thomsen_2014_Drug.Metab.Dispos_42_126
PubMedID: 24141856
Gene_locus related to this paper: human-CES1

Title : Carboxylesterase 1 gene duplication and mRNA expression in adipose tissue are linked to obesity and metabolic function - Friedrichsen_2013_PLoS.One_8_e56861
Author(s) : Friedrichsen M , Poulsen P , Wojtaszewski J , Hansen PR , Vaag A , Rasmussen HB
Ref : PLoS ONE , 8 :e56861 , 2013
Abstract : CONTEXT AND AIMS: Carboxylesterase 1 (CES1) appears to play an important role in the control of the metabolism of triglycerides and cholesterol in adipocytes and other cell types including hepatocytes. Therefore, it is relevant to gain insights into the genetic versus non-genetic mechanisms involved in the control of mRNA expression. Here, we investigated mRNA expression level in adipose tissue and its association with measures of adiposity and metabolic function in a population of elderly twins. Furthermore, the heritability of mRNA expression level in adipose tissue and the effect of gene duplication were assessed. METHODOLOGY: A total of 295 monozygotic and dizygotic twin subjects (62-83 years) with ( = 48) or without ( = 247) type 2 diabetes mellitus were enrolled in the study. They were subjected to a standard oral glucose tolerance test and excision of abdominal subcutaneous fat biopsies during the fasting state. Levels of mRNA and copy number of the gene were assessed by quantitative PCR.
RESULTS: mRNA expression level in adipose tissue was positively associated with body-mass index (<0.001), homeostasis model assessment-insulin resistance ( = 0.003) and level of fasting glucose ( = 0.002), insulin ( = 0.006), and triglycerides ( = 0.003). The heritability for the expression of mRNA in adipose tissue was high. gene duplication was positively associated with insulin sensitivity ( = 0.05) as well as glucose tolerance ( = 0.03) and negatively associated with homeostasis model assessment-insulin resistance ( = 0.02). Duplication of was not linked to mRNA level of this gene ( = 0.63). CONCLUSION: CES1 mRNA in adipose tissue appears to be under strong genetic control and was associated with measures of metabolic function raising the possibility of a potential role of this enzyme in the development of type 2 diabetes mellitus. Further studies are needed to understand the potential effect of gene duplication on adipocyte and whole-body metabolic functions.
ESTHER : Friedrichsen_2013_PLoS.One_8_e56861
PubMedSearch : Friedrichsen_2013_PLoS.One_8_e56861
PubMedID: 23468884
Gene_locus related to this paper: human-CES1

Title : High level expression, purification and activation of human dipeptidyl peptidase I from mammalian cells - Yang_2011_Protein.Expr.Purif_76_59
Author(s) : Yang W , Xia W , Mao J , Xu D , Chen J , Feng S , Wang J , Li H , Theisen CF , Petersen JM , Thorolfsson M , Rasmussen HB , Junker F , Boel E , Su J
Ref : Protein Expr Purif , 76 :59 , 2011
Abstract : Dipeptidyl peptidase I (DPPI) plays a crucial role in maturation of many regulatory peptides and has been suggested as a pharmaceutical target in several inflammatory diseases. It is also a useful processing enzyme for the generation of authentic protein products by catalyzing the removal of N-terminal fusion peptides. We used a robust transient transfection system in human embryonic kidney 293 cells to exploit expression and activation of DPPI from chicken, rat and man for the development of an industrial production process. The expression of human and rat DPPI was significantly higher in the human HEK293 cell line than that obtained with avian DPPI. A CHO K1SV stable cell line was selected as the optimal stable host system for production of human DPPI yielding expression levels higher than 1.5 g/L. The secreted pro-DPPI underwent auto-maturation during defined buffer conditions during the purification steps. Active human DPPI was purified with a three-step purification strategy employing: Butyl Sepharose 4 Fast Flow, Sephadex G-25 Medium and Q Sepharose Fast Flow chromatography. The final yield of active enzyme was approximately 1 g/L cell culture. The enzyme exhibited exopeptidase activity against both a dipeptide-p-nitroanilide substrate and N-terminally extended MEAE-hGH (Met-Glu-Ala-Glu-human growth hormone). In conclusion, an efficient production process for recombinant human DPPI has been developed including a highly efficient and stable CHO cell system and an efficient purification procedure, which is simple and easy to scale for industrial purposes. The present data facilitates not only industrial applications of DPPI as a processing enzyme, but also provides active enzyme useful in the identification of small molecule inhibitors.
ESTHER : Yang_2011_Protein.Expr.Purif_76_59
PubMedSearch : Yang_2011_Protein.Expr.Purif_76_59
PubMedID: 20828618

Title : Common variants conferring risk of schizophrenia - Stefansson_2009_Nature_460_744
Author(s) : Stefansson H , Ophoff RA , Steinberg S , Andreassen OA , Cichon S , Rujescu D , Werge T , Pietilainen OP , Mors O , Mortensen PB , Sigurdsson E , Gustafsson O , Nyegaard M , Tuulio-Henriksson A , Ingason A , Hansen T , Suvisaari J , Lonnqvist J , Paunio T , Borglum AD , Hartmann A , Fink-Jensen A , Nordentoft M , Hougaard D , Norgaard-Pedersen B , Bottcher Y , Olesen J , Breuer R , Moller HJ , Giegling I , Rasmussen HB , Timm S , Mattheisen M , Bitter I , Rethelyi JM , Magnusdottir BB , Sigmundsson T , Olason P , Masson G , Gulcher JR , Haraldsson M , Fossdal R , Thorgeirsson TE , Thorsteinsdottir U , Ruggeri M , Tosato S , Franke B , Strengman E , Kiemeney LA , Melle I , Djurovic S , Abramova L , Kaleda V , Sanjuan J , de Frutos R , Bramon E , Vassos E , Fraser G , Ettinger U , Picchioni M , Walker N , Toulopoulou T , Need AC , Ge D , Yoon JL , Shianna KV , Freimer NB , Cantor RM , Murray R , Kong A , Golimbet V , Carracedo A , Arango C , Costas J , Jonsson EG , Terenius L , Agartz I , Petursson H , Nothen MM , Rietschel M , Matthews PM , Muglia P , Peltonen L , St Clair D , Goldstein DB , Stefansson K , Collier DA
Ref : Nature , 460 :744 , 2009
Abstract : Schizophrenia is a complex disorder, caused by both genetic and environmental factors and their interactions. Research on pathogenesis has traditionally focused on neurotransmitter systems in the brain, particularly those involving dopamine. Schizophrenia has been considered a separate disease for over a century, but in the absence of clear biological markers, diagnosis has historically been based on signs and symptoms. A fundamental message emerging from genome-wide association studies of copy number variations (CNVs) associated with the disease is that its genetic basis does not necessarily conform to classical nosological disease boundaries. Certain CNVs confer not only high relative risk of schizophrenia but also of other psychiatric disorders. The structural variations associated with schizophrenia can involve several genes and the phenotypic syndromes, or the 'genomic disorders', have not yet been characterized. Single nucleotide polymorphism (SNP)-based genome-wide association studies with the potential to implicate individual genes in complex diseases may reveal underlying biological pathways. Here we combined SNP data from several large genome-wide scans and followed up the most significant association signals. We found significant association with several markers spanning the major histocompatibility complex (MHC) region on chromosome 6p21.3-22.1, a marker located upstream of the neurogranin gene (NRGN) on 11q24.2 and a marker in intron four of transcription factor 4 (TCF4) on 18q21.2. Our findings implicating the MHC region are consistent with an immune component to schizophrenia risk, whereas the association with NRGN and TCF4 points to perturbation of pathways involved in brain development, memory and cognition.
ESTHER : Stefansson_2009_Nature_460_744
PubMedSearch : Stefansson_2009_Nature_460_744
PubMedID: 19571808

Title : Selectivity among dipeptidyl peptidases of the S9b family - Bjelke_2006_Cell.Mol.Biol.(Noisy-le-grand)_52_3
Author(s) : Bjelke JR , Kanstrup AB , Rasmussen HB
Ref : Cellular & Molecular Biology (Noisy-le-grand) , 52 :3 , 2006
Abstract : Dipeptidyl peptidase IV is a serine protease with an indirect role in antihyperglycaemia via degradation of the incretin hormones glucagon-like peptide 1 and glucose-dependent insulinotropic polypeptide. Inhibition of the DPP-IV is thus a potential therapeutic strategy for type 2 diabetes. In this study, we have investigated upon selectivity of dipeptidyl peptidase IV compared to two other members of the S9b family, dipeptidyl peptidase 8 and 9, based on kinetic analyses of the pancreatic peptide hormones neuropeptide Y and peptide YY. We report a striking 250-fold preference for cleavage of neuropeptide Y compared to peptide YY observed for DPP-8/-9, but not for DPP-IV. This difference appears to be linked to differences in the S1' pocket within the active site, particularly via flexibility of the oxyanion stabilizing residue Y547. These aspects are discussed in relation to available protein structures of DPP-IV and data on DPP-IV selective inhibitors.
ESTHER : Bjelke_2006_Cell.Mol.Biol.(Noisy-le-grand)_52_3
PubMedSearch : Bjelke_2006_Cell.Mol.Biol.(Noisy-le-grand)_52_3
PubMedID: 17543192

Title : Dipeptidyl peptidases 8 and 9: specificity and molecular characterization compared with dipeptidyl peptidase IV - Bjelke_2006_Biochem.J_396_391
Author(s) : Bjelke JR , Christensen J , Nielsen PF , Branner S , Kanstrup AB , Wagtmann N , Rasmussen HB
Ref : Biochemical Journal , 396 :391 , 2006
Abstract : Dipeptidyl peptidases 8 and 9 have been identified as gene members of the S9b family of dipeptidyl peptidases. In the present paper, we report the characterization of recombinant dipeptidyl peptidases 8 and 9 using the baculovirus expression system. We have found that only the full-length variants of the two proteins can be expressed as active peptidases, which are 882 and 892 amino acids in length for dipeptidyl peptidase 8 and 9 respectively. We show further that the purified proteins are active dimers and that they show similar Michaelis-Menten kinetics and substrate specificity. Both cleave the peptide hormones glucagon-like peptide-1, glucagon-like peptide-2, neuropeptide Y and peptide YY with marked kinetic differences compared with dipeptidyl peptidase IV. Inhibition of dipeptidyl peptidases IV, 8 and 9 using the well-known dipeptidyl peptidase IV inhibitor valine pyrrolidide resulted in similar K(i) values, indicating that this inhibitor is non-selective for any of the three dipeptidyl peptidases.
ESTHER : Bjelke_2006_Biochem.J_396_391
PubMedSearch : Bjelke_2006_Biochem.J_396_391
PubMedID: 16475979
Gene_locus related to this paper: human-DPP9

Title : Tyrosine 547 constitutes an essential part of the catalytic mechanism of dipeptidyl peptidase IV - Bjelke_2004_J.Biol.Chem_279_34691
Author(s) : Bjelke JR , Christensen J , Branner S , Wagtmann N , Olsen C , Kanstrup AB , Rasmussen HB
Ref : Journal of Biological Chemistry , 279 :34691 , 2004
Abstract : Human dipeptidyl peptidase IV (DPP-IV) is a ubiquitously expressed type II transmembrane serine protease. It cleaves the penultimate positioned prolyl bonds at the N terminus of physiologically important peptides such as the incretin hormones glucagon-like peptide 1 and glucose-dependent insulinotropic peptide. In this study, we have characterized different active site mutants. The Y547F mutant as well as the catalytic triad mutants S630A, D708A, and H740L showed less than 1% wild type activity. X-ray crystal structure analysis of the Y547F mutant revealed no overall changes compared with wild type apoDPP-IV, except the ablation of the hydroxyl group of Tyr(547) and a water molecule positioned in close proximity to Tyr(547). To elucidate further the reaction mechanism, we determined the crystal structure of DPP-IV in complex with diisopropyl fluorophosphate, mimicking the tetrahedral intermediate. The kinetic and structural findings of the tyrosine residue are discussed in relation to the catalytic mechanism of DPP-IV and to the inhibitory mechanism of the 2-cyanopyrrolidine class of potent DPP-IV inhibitors, proposing an explanation for the specificity of this class of inhibitors for the S9b family among serine proteases.
ESTHER : Bjelke_2004_J.Biol.Chem_279_34691
PubMedSearch : Bjelke_2004_J.Biol.Chem_279_34691
PubMedID: 15175333
Gene_locus related to this paper: human-DPP4

Title : Crystal structure of human dipeptidyl peptidase IV\/CD26 in complex with a substrate analog - Rasmussen_2003_Nat.Struct.Biol_10_19
Author(s) : Rasmussen HB , Branner S , Wiberg FC , Wagtmann N
Ref : Nat Struct Biol , 10 :19 , 2003
Abstract : Dipeptidyl peptidase IV (DPP-IV/CD26) is a multifunctional type II transmembrane serine peptidase. This enzyme contributes to the regulation of various physiological processes, including blood sugar homeostasis, by cleaving peptide hormones, chemokines and neuropeptides. We have determined the 2.5 A structure of the extracellular region of DPP-IV in complex with the inhibitor valine-pyrrolidide. The catalytic site is located in a large cavity formed between the alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. Both domains participate in inhibitor binding. The structure indicates how substrate specificity is achieved and reveals a new and unexpected opening to the active site.
ESTHER : Rasmussen_2003_Nat.Struct.Biol_10_19
PubMedSearch : Rasmussen_2003_Nat.Struct.Biol_10_19
PubMedID: 12483204
Gene_locus related to this paper: human-DPP4