Involvement of Human Blood Arylesterases and Liver Microsomal Carboxylesterases in Nafamostat Hydrolysis

Yamaori, Satoshi; Fujiyama, Nobuhiro; Kushihara, Mika; Funahashi, Tatsuya; Kimura, Toshiyuki; Yamamoto, Ikuo; Sone, Tomomichi; Isobe, Masakazu; Ohshima, Tohru; Matsumura, Kenji; Oda, M.; Watanabe, Kazuhito

doi:10.2133/dmpk.21.147

Cited by 62 publications

(60 citation statements)

References 30 publications

(24 reference statements)

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“…Finally, the hydrolysis reaction was measured in HLM in the presence of various known chemical inhibitors of hydrolyzing enzymes. DTNB, BW284c51, and ethopropazine are specific inhibitors of arylesterase, acetylcholine esterase, and butyrylcholinesterase, respectively (Yamaori et al, 2006) and had no inhibitory effect on CDP323 hydrolysis (Fig. 8).…”

Section: Hydrolysis and Transesterification Of Cdp323 Prodrug 157mentioning

confidence: 97%

In Vitro Hydrolysis and Transesterification of CDP323, an α4β1/α4β7 Integrin Antagonist Ester Prodrug

et al. 2013

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We identified the enzyme(s) involved in the hydrolysis of the ethyl ester prodrug CDP323 (C 28 H 29 BrN 4 0 3 ) and characterized its transesterification in the presence of ethanol with special emphasis on the risks of drug-drug interaction. The hydrolysis of CDP323 was evaluated in vitro using human liver and intestinal microsomes and recombinant human carboxylesterases (hCES1 and 2) and was shown to be approximately 20-fold higher in human liver microsomes when compared with human intestinal microsomes and in hCES1 when compared with hCES2. Nonspecific inhibitors of carboxylesterases significantly inhibited the hydrolysis of CDP323 (>80% inhibition) while specific inhibitors of CES2, acetylcholine esterase, arylesterase, and butyrylcholinesterase did not impair the hydrolysis reaction. The effect of ethanol on the kinetic parameters for hydrolysis was investigated, demonstrating that at high concentration (2%), Michaelis-Menten constant (K m ), maximum velocity (V max ), and intrinsic clearance (CL int ) for the formation of the hydrolyzed product were decreased (∼40%). The use of deuterated ethanol allowed more mechanistic investigations of the transesterification mechanism and showed that the intrinsic clearance based on parent loss was not impaired in the presence of alcohol. Overall, our data demonstrate that CDP323 is mainly hydrolyzed by hCES1 and is prone to transesterification in the presence of ethanol. Transesterification mechanisms compete with hydrolysis without impairing the overall clearance of the ester prodrug. Based on in vitro results, the risk of a clinically significant drug-drug interaction with ethanol is anticipated to be low.

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Section: Hydrolysis and Transesterification Of Cdp323 Prodrug 157mentioning

confidence: 97%

In Vitro Hydrolysis and Transesterification of CDP323, an α4β1/α4β7 Integrin Antagonist Ester Prodrug

et al. 2013

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“…The variability in GB hydrolase activity of human plasma was smaller than those in butyrylcholinesterase activity (2.0-fold) (Table 1), aspirin hydrolase activity (2.2-fold), 14) and fluazifop-butyl hydrolase activity (approximately 4-fold), 11) although the variation in plasma GB hydrolase activity was larger than that in plasma nafamostat hydrolase activity (1.2-fold). 10) Interindividual difference in erythrocyte activity of the GB hydrolysis was slightly smaller than those in acetylcholinesterase activity (1.3-fold) ( Table 1) and nafamostat hydrolase activity (1.4-fold) reported previously. 10) Thus, GB hydrolytic activity of human blood fractions may show relatively smaller interindividual variation.…”

Section: Discussionmentioning

confidence: 45%

“…10) Interindividual difference in erythrocyte activity of the GB hydrolysis was slightly smaller than those in acetylcholinesterase activity (1.3-fold) ( Table 1) and nafamostat hydrolase activity (1.4-fold) reported previously. 10) Thus, GB hydrolytic activity of human blood fractions may show relatively smaller interindividual variation.…”

Section: Discussionmentioning

confidence: 45%

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Butyrylcholinesterase and Erythrocyte Sulfhydryl-dependent Enzyme Hydrolyze Gabexate in Human Blood

Yamaori

Kushihara

Fujiyama

et al. 2007

JOURNAL OF HEALTH SCIENCE

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Gabexate (GB), a serine protease inhibitor that is widely used for the treatment of acute pancreatitis and disseminated intravascular coagulation, has been reported to be partly hydrolyzed by human serum albumin. However, other enzymes responsible for GB hydrolysis in human blood remain unclear. In this study, we examined in vitro metabolism of GB with human blood samples. The hydrolytic activities of the plasma and erythrocytes at 100 µM of GB were 167 ± 26 and 151 ± 9 nmol/min/ml blood fraction (mean ± S.D., n = 8), respectively. Kinetic analysis indicated that V max and K m values were 1.75 µmol/min/ml blood fraction and 529 µM for the plasma and 10.6 µmol/min/ml blood fraction and 7360 µM for the erythrocytes, respectively. The activity of human plasma was inhibited by ethopropazine, a butyrylcholinesterase inhibitor (27% inhibition at 100 µM). Furthermore, the plasma activity was inhibited by 5,5 -dithiobis(2-nitrobenzoic acid) (DTNB) (40% inhibition at 5000 µM), suggesting the involvement of albumin in the plasma GB hydrolysis. The erythrocyte activity was also decreased by DTNB (56% inhibition at 5000 µM), implying that this activity was dependent on the presence of sulfhydryl group(s), while little or no inhibition of the activity was seen with phenylmethylsulfonyl fluoride, diisopropyl fluorophosphate, and BW284C51. Butyrylcholinesterase from human serum showed GB hydrolytic activity with V max of 363 nmol/min/mg protein and K m of 263 µM. These results suggest that, in addition to albumin, butyrylcholinesterase and erythrocyte sulfhydryl-dependent enzyme are responsible for GB hydrolysis in human blood.

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“…The activity at 20 M in HLM was not inhibited by 10 M ethopropazine, an inhibitor of butyrylcholinesterase (Yamaori et al, 2006), and was not activated by 1 mM CaCl 2 , which is required for exerting paraoxonase activity (Kuo and La Du, 1998) (data not shown). Thus, we first found the involvement of human CES2 in the flutamide hydrolysis.…”

Section: Resultsmentioning

confidence: 94%

Contributions of Arylacetamide Deacetylase and Carboxylesterase 2 to Flutamide Hydrolysis in Human Liver

Kobayashi

Fukami²,

Shimizu³

et al. 2012

Drug Metab Dispos

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ABSTRACT:Flutamide, an antiandrogen drug, is widely used for the treatment of prostate cancer. The major metabolic pathways of flutamide are hydroxylation and hydrolysis. The hydrolyzed metabolite, 5-amino-2-nitrobenzotrifluoride (FLU-1), is further metabolized to N-hydroxy FLU-1, an assumed hepatotoxicant. Our previous study demonstrated that arylacetamide deacetylase (AADAC), one of the major serine esterases expressed in the human liver and gastrointestinal tract, catalyzes the flutamide hydrolysis. However, the enzyme kinetics in human tissue microsomes were not consistent with the kinetics by recombinant human AADAC. Thus, it seemed that AADAC is not the sole enzyme responsible for flutamide hydrolysis in human. In the present study, we found that recombinant carboxylesterase (CES) 2 could hydrolyze flutamide at low concentrations of flutamide. In the inhibition assay, the flutamide hydrolase activities at a flutamide concentration of 5 M in human liver and jejunum microsomes were strongly inhibited by a selective CES2 inhibitor, 10 M loperamide, with the residual activities of 22.9 ؎ 3.5 and 18.6 ؎ 0.7%, respectively. These results suggest that CES2 is also involved in the flutamide hydrolysis in human tissues. Using six individual human livers, the contributions of AADAC and CES2 to flutamide hydrolysis were estimated by using the relative activity factor. The relative contribution of CES2 was approximately 75 to 99% at the concentration of 5 M flutamide. In contrast, the relative contribution of AADAC increased in parallel with the concentration of flutamide. Thus, CES2, rather than AA-DAC, largely contributed to the flutamide hydrolysis in clinical therapeutics.

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Involvement of Human Blood Arylesterases and Liver Microsomal Carboxylesterases in Nafamostat Hydrolysis

Cited by 62 publications

References 30 publications

In Vitro Hydrolysis and Transesterification of CDP323, an α4β1/α4β7 Integrin Antagonist Ester Prodrug

In Vitro Hydrolysis and Transesterification of CDP323, an α4β1/α4β7 Integrin Antagonist Ester Prodrug

Butyrylcholinesterase and Erythrocyte Sulfhydryl-dependent Enzyme Hydrolyze Gabexate in Human Blood

Contributions of Arylacetamide Deacetylase and Carboxylesterase 2 to Flutamide Hydrolysis in Human Liver

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