Uehara S

References (8)

Title : Drug-oxidizing and conjugating non-cytochrome P450 (non-P450) enzymes in cynomolgus monkeys and common marmosets as preclinical models for humans - Uno_2021_Biochem.Pharmacol_197_114887
Author(s) : Uno Y , Uehara S , Yamazaki H
Ref : Biochemical Pharmacology , 197 :114887 , 2021
Abstract : Many drug oxidations and conjugations are mediated by a variety of cytochromes P450 (P450) and non-P450 enzymes in humans and non-human primates. These non-P450 enzymes include aldehyde oxidases (AOX), carboxylesterases (CES), flavin-containing monooxygenases (FMO), glutathione S-transferases (GST), arylamine N-acetyltransferases (NAT),sulfotransferases (SULT), and uridine 5'-diphospho-glucuronosyltransferases (UGT) and their substrates include both endobiotics and xenobiotics. Cynomolgus macaques (Macaca fascicularis, an Old-World monkey) are widely used in preclinical studies because of their genetic and physiological similarities to humans. However, many reports have indicated the usefulness of common marmosets (Callithrix jacchus, a New World monkey) as an alternative non-human primate model. Although knowledge of the drug-metabolizing properties of non-P450 enzymes in non-human primates is relatively limited, new research has started to provide an insight into the molecular characteristics of these enzymes in cynomolgus macaques and common marmosets. This mini-review provides collective information on the isoforms of non-P450 enzymes AOX, CES, FMO, GST, NAT, SULT, and UGT and their enzymatic profiles in cynomolgus macaques and common marmosets. In general, these non-P450 cynomolgus macaque and marmoset enzymes have high sequence identities and similar substrate recognitions to their human counterparts. However, these enzymes also exhibit some limited differences in function between species, just as P450 enzymes do, possibly due to small structural differences in amino acid residues. The findings summarized here provide a foundation for understanding the molecular mechanisms of polymorphic non-P450 enzymes and should contribute to the successful application of non-human primates as model animals for humans.
ESTHER : Uno_2021_Biochem.Pharmacol_197_114887
PubMedSearch : Uno_2021_Biochem.Pharmacol_197_114887
PubMedID: 34968483

Title : Differences in hydrolase activities in the liver and small intestine between marmosets and humans - Honda_2021_Drug.Metab.Dispos__
Author(s) : Honda S , Fukami T , Hirosawa K , Tsujiguchi T , Zhang Y , Nakano M , Uehara S , Uno Y , Yamazaki H , Nakajima M
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , : , 2021
Abstract : 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 cynomolgusmonkeys. 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 dueto 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. Significance Statement This study confirmed that there are large differences in hydrolase activities between humans and marmosets by characterizing marmoset hydrolase activities for compounds that are substrates of human CES1, CES2, or AADAC. The data obtained in this study may be useful for considering whether marmosets are appropriate for examining the pharmacokinetics and efficacies of new chemical entities in preclinical studies.
ESTHER : Honda_2021_Drug.Metab.Dispos__
PubMedSearch : Honda_2021_Drug.Metab.Dispos__
PubMedID: 34135089

Title : Molecular characterization and polymorphisms of butyrylcholinesterase in cynomolgus macaques - Uno_2018_J.Med.Primatol_47_185
Author(s) : Uno Y , Uehara S , Mahadhi HMD , Ohura K , Hosokawa M , Imai T
Ref : J Med Primatol , 47 :185 , 2018
Abstract : BACKGROUND: Butyrylcholinesterase (BChE), an enzyme essential for drug metabolism, has been investigated as antidotes against organophosphorus nerve agents, and the efficacy and safety have been studied in cynomolgus macaques. BChE polymorphisms partly account for variable BChE activities among individuals in humans, but have not been investigated in cynomolgus macaques. METHODS: Molecular characterization was carried out by analyzing primary sequence, gene, tissue expression, and genetic variants. RESULTS: In cynomolgus and human BChE, phylogenetically closely related, amino acid residues important for enzyme function were conserved, and gene and genomic structure were similar. Cynomolgus BChE mRNA was most abundantly expressed in liver among the 10 tissue types analyzed. Re-sequencing found 26 non-synonymous genetic variants in 121 cynomolgus and 23 rhesus macaques, indicating that macaque BChE is polymorphic, although none of these variants corresponded to the null or defective alleles of human BChE. CONCLUSIONS: These results suggest molecular similarities of cynomolgus and human BChE.
ESTHER : Uno_2018_J.Med.Primatol_47_185
PubMedSearch : Uno_2018_J.Med.Primatol_47_185
PubMedID: 29573432
Gene_locus related to this paper: macfa-BCHE

Title : Differences in Intestinal Hydrolytic Activities between Cynomolgus Monkeys and Humans: Evaluation of Substrate Specificities Using Recombinant Carboxylesterase 2 Isozymes - Igawa_2016_Mol.Pharm_13_3176
Author(s) : Igawa Y , Fujiwara S , Ohura K , Hirokawa T , Nishizawa Y , Uehara S , Uno Y , Imai T
Ref : Mol Pharm , 13 :3176 , 2016
Abstract : Cynomolgus monkeys, used as an animal model to predict human pharmacokinetics, occasionally show different oral absorption patterns to humans due to differences in their intestinal metabolism. In this study, we investigated the differences between intestinal hydrolytic activities in cynomolgus monkeys and humans, in particular the catalyzing activities of their carboxylesterase 2 (CES2) isozymes. For this purpose we used both human and monkey microsomes and recombinant enzymes derived from a cell culture system. Monkey intestinal microsomes showed lower hydrolytic activity than human microsomes for several substrates. Interestingly, in contrast to human intestinal hydrolysis, which is not enantioselective, monkey intestine showed preferential R-form hydrolysis of propranolol derivatives. Recombinant CES2 isozymes from both species, mfCES2v3 from monkeys and human hCE2, showed similar metabolic properties to their intestinal microsomes when expressed in HEK293 cells. Recombinant hCE2 and mfCES2v3 showed similar Km values for both enantiomers of all propranolol derivatives tested. However, recombinant mfCES2v3 showed extreme R-enantioselective hydrolysis, and both hCE2 and mfCES2v3 showed lower activity for O-3-methyl-n-butyryl propranolol than for O-n-valeryl and O-2-methyl-n-butyryl propranolol. This lower hydrolytic activity was characterized by lower Vmax values. Docking simulations of the protein-ligand complex demonstrated that the enantioselectivity of mfCES2v3 for propranolol derivatives was possibly caused by the orientation of its active site being deformed by an amino acid change of Leu107 to Gln107 and the insertion of Met309, compared with hCE2. In addition, molecular dynamics simulation indicated the possibility that the interatomic distance between the catalytic triad and the substrate was elongated by a 3-positioned methyl in the propranolol derivatives. Overall, these findings will help us to understand the differences in intestinal hydrolytic activities between cynomolgus monkeys and humans.
ESTHER : Igawa_2016_Mol.Pharm_13_3176
PubMedSearch : Igawa_2016_Mol.Pharm_13_3176
PubMedID: 27454346

Title : Serum butyrylcholinesterase and the risk of future type 2 diabetes: the Kansai Healthcare Study - Sato_2014_Clin.Endocrinol.(Oxf)_80_362
Author(s) : Sato KK , Hayashi T , Maeda I , Koh H , Harita N , Uehara S , Onishi Y , Oue K , Nakamura Y , Endo G , Kambe H , Fukuda K
Ref : Clinical Endocrinology (Oxf) , 80 :362 , 2014
Abstract : OBJECTIVE: Butyrylcholinesterase is synthesized in the liver. The serum butyrylcholinesterase level has been cross-sectionally reported to be higher in patients with diabetes, hyperlipidaemia, obesity and fatty liver than in those without them. It is not known whether serum butyrylcholinesterase is associated with the risk of future type 2 diabetes. DESIGN: A prospective cohort study. PARTICIPANTS: A total of 8470 Japanese men aged 40-55 years without type 2 diabetes at baseline. MEASUREMENTS: Type 2 diabetes was diagnosed if a fasting plasma glucose (FPG) level was >/=7.0 mmol/l, if a HbA1 c level was >/=6.5% or if participants were taking oral hypoglycaemic medication or insulin.
RESULTS: During the 42 227 person-years of follow-up, 868 cases had developed type 2 diabetes. Serum butyrylcholinesterase was significantly positively correlated with body mass index (BMI), FPG, alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT) and triglycerides (TG), whereas negatively with high-density lipoprotein (HDL) cholesterol. In Cox proportional hazards models, after adjusting for age, BMI, FPG, alcohol consumption, smoking habit, walk to work, regular leisure-time physical activity and family history of diabetes, the highest quartile (398-806 IU/l) of serum butyrylcholinesterase increased the risk of type 2 diabetes compared with the lowest quartile (56-311 IU/l) [hazard ratio (HR) 1.41 (95% confidence interval (CI), 1.14-1.74)]. After further adjusting for ALT and GGT, this association remained [HR 1.40 (95% CI, 1.13-1.73)]. Furthermore, this association was significant independent of TG and HDL cholesterol.
CONCLUSIONS: Elevated serum butyrylcholinesterase was independently associated with an increased risk of future type 2 diabetes.
ESTHER : Sato_2014_Clin.Endocrinol.(Oxf)_80_362
PubMedSearch : Sato_2014_Clin.Endocrinol.(Oxf)_80_362
PubMedID: 23418907

Title : Systematic identification and characterization of carboxylesterases in cynomolgus macaques - Uno_2014_Drug.Metab.Dispos_42_2002
Author(s) : Uno Y , Uehara S , Hosokawa M , Imai T
Ref : Drug Metabolism & Disposition: The Biological Fate of Chemicals , 42 :2002 , 2014
Abstract : Carboxylesterase (CES) is important for detoxification of a wide range of drugs and xenobiotics and catalyzes cholesterol and fatty acid metabolism. Cynomolgus macaques are widely used in drug metabolism studies; however, cynomolgus CES has not been fully investigated at molecular levels, partly due to the lack of gene information. In this study, we isolated and characterized cDNAs for CES homologous to human CES1, CES2, and CES5A in cynomolgus macaques. By genome analysis, in the cynomolgus macaque genome, three gene sequences were found for CES1(v1-3) and CES2(v1-3), whereas one gene sequence was found for CES5A. Cynomolgus CES1, CES2, and CES5A genes were located in the genomic regions corresponding to the human genes. We successfully identified CES1v1, CES1v2, CES2v1, CES2v3, and CES5A cDNAs from cynomolgus liver. Sequence analysis showed that amino acid sequences of each CES were highly homologous to that of the human homolog. All five CESs had sequences characteristic for CES enzymes, including the catalytic triad and oxyanion hole loop. By quantitative polymerase chain reaction, the most abundant expression of CES mRNAs among the 10 tissue types analyzed was observed in liver (CES1v1 and CES2v3 mRNAs), jejunum (CES2v1 mRNAs), and kidney (CES1v2 and CES5A mRNA), the organs important for drug metabolism and excretion. The results indicated that cynomolgus macaques express at least five CES genes, which potentially encode intact CES proteins.
ESTHER : Uno_2014_Drug.Metab.Dispos_42_2002
PubMedSearch : Uno_2014_Drug.Metab.Dispos_42_2002
PubMedID: 25256558
Gene_locus related to this paper: macfa-CES1v2 , macfa-CES1v1 , macfa-CES5 , macfa-CES2v3

Title : Identification of human cytochrome P450 isoforms and esterases involved in the metabolism of mirabegron, a potent and selective beta(3)-adrenoceptor agonist - Takusagawa_2012_Xenobiotica_42_957
Author(s) : Takusagawa S , Yajima K , Miyashita A , Uehara S , Iwatsubo T , Usui T
Ref : Xenobiotica , 42 :957 , 2012
Abstract : Human cytochrome P450 CYP enzymes and esterases involved in the metabolism of mirabegron a potent and selective human beta(3)-adrenoceptor agonist intended for the treatment of overactive bladder were identified in in vitro studies Incubations of mirabegron with recombinant human CYP enzymes showed significant metabolism of mirabegron by CYP2D6 and CYP3A4 only Correlation analyses showed a significant correlation between mirabegron metabolism and testosterone 6beta-hydroxylation CYP3A4/5 marker activity In inhibition studies using antiserum against CYP3A4 a strong inhibition at maximum 80 inhibition of the metabolism of mirabegron was observed whereas the inhibitory effects of monoclonal antibodies against CYP2D6 were small at maximum 10 inhibition These findings suggest that CYP3A4 is the primary CYP enzyme responsible for in vitro oxidative metabolism of mirabegron with a minor role of CYP2D6 Mirabegron hydrolysis was catalyzed in human blood plasma and butyrylcholinesterase BChE solution but not in human liver microsomes intestinal microsomes liver S9 intestinal S9 and recombinant acetylcholinesterase solution K(m values of mirabegron hydrolysis in human blood plasma and BChE solution were all similar 13.4-15.2 muM The inhibition profiles in human blood and plasma were also similar to those in BChE solution suggesting that mirabegron hydrolysis is catalyzed by BChE.
ESTHER : Takusagawa_2012_Xenobiotica_42_957
PubMedSearch : Takusagawa_2012_Xenobiotica_42_957
PubMedID: 22509825

Title : Studies on the therapeutic effect of 2-pyridine aldoxime methiodide (2-PAM) in mammals following organophosphorus compound-poisoning (report III): distribution and antidotal effect of 2-PAM in rats - Uehara_1993_J.Toxicol.Sci_18_265
Author(s) : Uehara S , Hiromori T , Isobe N , Suzuki T , Kato T , Miyamoto J
Ref : Journal of Toxicological Sciences , 18 :265 , 1993
Abstract : The metabolic fate of 2-PAM and its antidotal effect on organophosphorus compound poisoning in rats were studied. When 14C-2-PAM was administered intravenously, the amount of 14C reaching the brain was small. Following administration by intramedullary injection, 14C was present in high concentrations in the brain, and 72-90% of the 14C present in the brain corresponded to the unchanged form of 2-PAM. 2-PAM was rapidly excreted into the urine and feces following either intramedullary or intravenous administration. The half-life of 2-PAM in the brain following intramedullary administration was 1.52 hr. Intramedullary administration of 2-PAM to rats poisoned with fenitrothion or malathion enabled their survival and induced reactivation of brain cholinesterase.
ESTHER : Uehara_1993_J.Toxicol.Sci_18_265
PubMedSearch : Uehara_1993_J.Toxicol.Sci_18_265
PubMedID: 8295230