An Orphan Esterase ABHD10 Modulates Probenecid Acyl Glucuronidation in Human Liver

Abstract

Probenecid, a widely used uricosuric agent, is mainly metabolized to probenecid acyl glucuronide (PRAG), which is considered a causal substance of severe allergic or anaphylactoid reactions. PRAG can be hydrolyzed (deglucuronidated) to probenecid. The purpose of this study was to identify enzymes responsible for probenecid acyl glucuronidation and PRAG deglucuronidation in human livers and to examine the effect of deglucuronidation in PRAG formation. In human liver homogenates (HLHs), the intrinsic clearance (CL int ) of PRAG deglucuronidation was much greater (497-fold) than that of probenecid acyl glucuronidation. Evaluation of PRAG formation by recombinant UDP-glucuronosyltransferase (UGT) isoforms and an inhibition study using HLHs as an enzyme source demonstrated that multiple UGT isoforms, including UGT1A1, UGT1A9, and UGT2B7, catalyzed probenecid acyl glucuronidation. We found that recombinant a/b hydrolase domain containing 10 (ABHD10) substantially catalyzed PRAG deglucuronidation activity, whereas carboxylesterases did not. Similar inhibitory patterns by chemicals between HLHs and recombinant ABHD10 supported the major contribution of ABHD10 to PRAG deglucuronidation in human liver. Interestingly, it was demonstrated that the CL int value of probenecid acyl glucuronidation in HLHs was increased by 1.7-fold in the presence of phenylmethylsulfonyl fluoride, which potently inhibited ABHD10 activity. In conclusion, we found that PRAG deglucuronidation catalyzed by ABHD10 suppressively regulates PRAG formation via multiple UGT enzymes in human liver. The balance of activities by these enzymes is important for the formation of PRAG, which may be associated with the adverse reactions observed after probenecid administration.

Involvement of Reactive Metabolites of Diclofenac in Cytotoxicity in Sandwich-Cultured Rat Hepatocytes

“…tolmetin, probenecid, and DIC. 41,42 The intracellular concentrations of DIC-AG at 1, 3, and 6 hours after DIC treatment were approximately 1% those of DIC. We choose the DIC dose of 300 mM for subsequent experiments because the intracellular concentrations of DIC-AG and 4 0 OH-DIC were decreased at 1,000 mM DIC due to cytotoxicity by DIC itself.…”

Physiologically Based Pharmacokinetic Models of Probenecid and Furosemide to Predict Transporter Mediated Drug-Drug Interactions

“…Therefore, probenecid absorption was described by optimization of the passive transcellular intestinal permeability (optimized value: 3.97 • 10 −4 cm/min, lipophilicity based calculated value: 3.12 • 10 −6 cm/min). The uptake of probenecid into the kidney, which is its site of action and metabolism (39)(40)(41), was assumed to be mediated by OAT3. The parameters (K M , k cat ) to describe this transport were optimized during parameter identification (see Table S2.3.1).…”

Identification of key genes involved in type 2 diabetic islet dysfunction: a bioinformatics study

“…Further study about the relation between ADHRS2 and diabetes needs to be conducted. A study has reported that human ABHD10 is closely related to probenecid acyl glucuronidation in liver [27]. There is currently no report whether ABHD10 is involved in diabetes.…”