Characterization of Species Differences in Tissue Diltiazem Deacetylation Identifies Ces2a as a Rat-Specific Diltiazem Deacetylase

Kurokawa, Takaya; Fukami, Tatsuki; Nakajima, Miki

doi:10.1124/dmd.115.064089

Cited by 13 publications

(4 citation statements)

References 34 publications

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“…To further confirm the involved enzymes, AKBA was incubated with HLM and HIM in the presence or absence of 100 μM esterase inhibitor: HA, EDTA, iso-OMPA, BNPP, and LPA (39)(40)(41). The concentrations of AKBA were the approximate K m values for HLM (20 μM), HIM (15 μM), and CE2 (6 μM).…”

Section: Chemical Inhibition Studymentioning

confidence: 99%

Metabolic Profile of 3-Acetyl-11-Keto-β-Boswellic Acid and 11-Keto-β-Boswellic Acid in Human Preparations In Vitro, Species Differences, and Bioactivity Variation

Cui

Tian

Ning

et al. 2016

AAPS J

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3-Acetyl-11-keto-β-boswellic acid (AKBA) and 11-keto-β-boswellic acid (KBA) are widely used in the clinic as anti-inflammatory drugs. However, these drugs have the poor bioavailability, which may be caused by their extensive metabolism. In this study, we systemically characterized both phase I and II metabolism of AKBA and KBA in vitro. In total, four major metabolites were firstly biosynthesized and identified using 1D and 2D NMR spectroscopy. Among them, three metabolites were novel. The kinetic parameters (K m , V max , CL int, and K i ) were also analyzed systematically in various biological samples. Finally, the deacetylation of AKBA and hydroxylation of KBA were confirmed to be the major metabolic pathways based on their large CL int and the high amounts of KBA (46.7%) and hydroxylated KBA (50.8%) along with a low amount of AKBA (2.50%) in human primary hepatocytes. Carboxylesterase 2 (CE2) selectively catalyzed the deacetylation of AKBA to form KBA. Although CYP3A4, CYP3A5, and CYP3A7 catalyzed the metabolism of KBA, CYP3A4 played a predominant role in the hydroxylation reaction of KBA in human. Notably, deacetylation and regioselective hydroxylation exhibited considerable species differences. Deacetylation was only observed in human liver microsomes and primary human hepatocytes; 21- and 20-mono-hydroxylation of KBA were primarily observed in human, monkey, and dog; and 16- and 30-mono-hydroxylation were observed in other species. More importantly, all four mono-hydroxylation metabolites exhibited a moderate anti-inflammatory activity. The 21- and 20-hydroxylation metabolites inhibited the expression of iNOS, the LPS-induced activation of IkBα and p65 phosphorylation, and suppressed p65 nuclear translocation in RAW264.7 cells.

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Section: Chemical Inhibition Studymentioning

confidence: 99%

Metabolic Profile of 3-Acetyl-11-Keto-β-Boswellic Acid and 11-Keto-β-Boswellic Acid in Human Preparations In Vitro, Species Differences, and Bioactivity Variation

Cui

Tian

Ning

et al. 2016

AAPS J

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“…The Ces1d protein is the mouse ortholog of human CES1 protein in terms of both amino acid sequence homology and enzyme function . The Ces/CES isoforms are distributed in several mammalian tissues; for example, Ces1d/CES1 is abundant in liver, lung, and adipose in mice and humans. − Further, CES1 is expressed in human monocytes and macrophages, , including human alveolar macrophages, whereas Ces1d mRNA is detected in mouse alveolar macrophages but not in either thioglycolate (TG)-elicited peritoneal macrophages or bone marrow-derived macrophages (our unpublished data). Together, these findings suggest that Ces1d/CES1 enzymes have a role in innate immune responses, particularly in lung tissues.…”

mentioning

confidence: 97%

Carboxylesterase 1d Inactivation Augments Lung Inflammation in Mice

Szafran

Borazjani

Scheaffer

et al. 2022

ACS Pharmacol. Transl. Sci.

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Carboxylesterases are members of the serine hydrolase superfamily and metabolize drugs, pesticides, and lipids. Previous research showed that inhibition of carboxylesterase 1 (CES1) in human macrophages altered the immunomodulatory effects of lipid mediators called prostaglandin glyceryl esters, which are produced by cyclooxygenase-catalyzed oxygenation of the endocannabinoid 2-arachidonoylglycerol (2-AG). Ces1d − the mouse ortholog of human CES1 − is the most abundant Ces isoform in murine lung tissues and alveolar macrophages and a major target of organophosphate poisons. Monoacylglycerol lipase (Magl) is also expressed in murine lung and is the main enzyme responsible for 2-AG catabolism. Several metabolic benefits are observed in Ces1d −/− mice fed a high-fat diet; thus, we wondered whether pharmacological and genetic inactivation of Ces1d in vivo might also ameliorate the acute inflammatory response to lipopolysaccharide (LPS). C57BL/6 mice were treated with WWL229 (Ces1d inhibitor) or JZL184 (Magl inhibitor), followed 30 min later by either LPS or saline. Wild-type (WT) and Ces1d −/− mice were also administered LPS to determine the effect of Ces1d knockout. Mice were sacrificed at 6 and 24 h, and cytokines were assessed in serum, lung, liver, and adipose tissues. Lipid mediators were quantified in lung tissues, while activity-based protein profiling and enzyme assays determined the extent of lung serine hydrolase inactivation by the inhibitors. WWL229 was shown to augment LPS-induced lung inflammation in a female-specific manner, as measured by enhanced neutrophil infiltration and Il1b mRNA. The marked Ces inhibition in female lung by 4 h after drug treatment might explain this sex difference, although the degree of Ces inhibition in female and male lungs was similar at 6 h. In addition, induction of lung Il6 mRNA and prostaglandin E 2 by LPS was more pronounced in Ces1d −/− mice than in WT mice. Thus, WWL229 inhibited lung Ces1d activity and augmented the female lung innate immune response, an effect observed in part in Ces1d −/− mice and Ces1d/CES1-deficient murine and human macrophages. In contrast, JZL184 attenuated LPS-induced Il1b and Il6 mRNA levels in female lung, suggesting that Ces1d and Magl have opposing effects. Mapping the immunomodulatory molecules/pathways that are regulated by Ces1d in the context of lung inflammation will require further research.

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“…The final version may differ from this version. (Kurokawa et al, 2015). (2) In dogs, the CES2 protein is not functional due to its instability (Yoshida et al, 2017), although CES2 mRNA is substantially expressed in the liver (Taketani et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Differences in Hydrolase Activities in the Liver and Small Intestine between Marmosets and Humans

et al. 2021

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For drug development, species differences in drug-metabolism reactions present obstacles for predicting pharmacokinetics in humans. We characterized the species differences in hydrolases among humans and mice, rats, dogs, and cynomolgus monkeys. In this study, to expand the series of such studies, we attempted to characterize marmoset hydrolases. We measured hydrolase activities for 24 compounds using marmoset liver and intestinal microsomes, as well as recombinant marmoset carboxylesterase (CES) 1, CES2, and arylacetamide deacetylase (AADAC). The contributions of CES1, CES2, and AADAC to hydrolysis in marmoset liver microsomes were estimated by correcting the activities by using the ratios of hydrolase protein levels in the liver microsomes and those in recombinant systems. For 6 out of 8 human CES1 substrates, the activities in marmoset liver microsomes were lower than those in human liver microsomes. For 2 human CES2 substrates and 3 out of 7 human AADAC substrates, the activities in marmoset liver microsomes were higher than those in human liver microsomes. Notably, among the 3 rifamycins, only rifabutin was hydrolyzed by marmoset tissue microsomes and recombinant AADAC. The activities for all substrates in marmoset intestinal microsomes tended to be lower than those in liver microsomes, which suggests that the first-pass effects of the CES and AADAC substrates are due to hepatic hydrolysis. In most cases, the sums of the values of the contributions of CES1, CES2, and AADAC were below 100%, which indicated the involvement of other hydrolases in marmosets. In conclusion, we clarified the substrate preferences of hydrolases in marmosets.

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Characterization of Species Differences in Tissue Diltiazem Deacetylation Identifies Ces2a as a Rat-Specific Diltiazem Deacetylase

Cited by 13 publications

References 34 publications

Metabolic Profile of 3-Acetyl-11-Keto-β-Boswellic Acid and 11-Keto-β-Boswellic Acid in Human Preparations In Vitro, Species Differences, and Bioactivity Variation

Metabolic Profile of 3-Acetyl-11-Keto-β-Boswellic Acid and 11-Keto-β-Boswellic Acid in Human Preparations In Vitro, Species Differences, and Bioactivity Variation

Carboxylesterase 1d Inactivation Augments Lung Inflammation in Mice

Differences in Hydrolase Activities in the Liver and Small Intestine between Marmosets and Humans

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