Glucuronidation of Edaravone by Human Liver and Kidney Microsomes: Biphasic Kinetics and Identification of UGT1A9 as the Major UDP-Glucuronosyltransferase Isoform

Edaravone was launched in Japan in 2001 and was the first neuroprotectant developed for the treatment of acute cerebral infarction. Edaravone is mainly eliminated as glucuronide conjugate in human urine (approximately 70%), but the mechanism involved in the elimination pathway remains unidentified. We investigated the glucuronidation of edaravone in human liver microsomes (HLM) and human kidney microsomes (HKM) and identified the major hepatic and renal UDP-glucuronosyltransferases (UGTs) involved. As we observed, edaravone glucuronidation in HLM and HKM exhibited biphasic kinetics. The intrinsic clearance of glucuronidation at high-affinity phase (CLint1) and low-affinity phase (CLint2) were 8.4 ± 3.3 and 1.3 ± 0.2 μl · min−1 · mg−1, respectively, for HLM and were 45.3 ± 8.2 and 1.8 ± 0.1 μl · min−1 · mg−1, respectively, for HKM. However, in microsomal incubations contained with 2% bovine serum albumin, CLint1 and CLint2 were 16.4 ± 1.2 and 3.7 ± 0.3 μl · min−1 · mg−1, respectively, for HLM and were 78.5 ± 3.9 and 3.6 ± 0.5 μl · min−1 · mg−1, respectively, for HKM. Screening with 12 recombinant UGTs indicated that eight UGTs (UGT1A1, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B7, and UGT2B17) produced a significant amount of glucuronide metabolite. Thus, six UGTs (UGT1A1, UGT1A6, UGT1A7, UGT1A9, UGT2B7, and UGT2B17) expressed in human liver or kidney were selected for kinetic studies. Among them, UGT1A9 exhibited the highest activity (CLint1 = 42.4 ± 9.5 μl · min−1 · mg−1), followed by UGT2B17 (CLint = 3.3 ± 0.4 μl · min−1 · mg−1) and UGT1A7 (CLint = 1.7 ± 0.2 μl · min−1 · mg−1). Inhibition study found that inhibitor of UGT1A9 (propofol) attenuated edaravone glucuronidation in HLM and HKM. In addition, edaravone glucuronidation in a panel of seven HLM was significantly correlated (r = 0.9340, p = 0.0021) with propofol glucuronidation. Results indicated that UGT1A9 was the main UGT isoform involved in edaravone glucuronidation in HLM and HKM.

[1]  P. Lapchak.  A critical assessment of edaravone acute ischemic stroke efficacy trials: is edaravone an effective neuroprotective therapy? , 2010, Expert opinion on pharmacotherapy.

[2]  K. Knights,et al.  Renal UDP-glucuronosyltransferases and the glucuronidation of xenobiotics and endogenous mediators , 2010, Drug metabolism reviews.

[3]  M. L. Maestro,et al.  The prevalence of uridine diphosphate-glucuronosyltransferase 1A9 (UGT1A9) gene promoter region single-nucleotide polymorphisms T-275A and C-2152T and its influence on mycophenolic acid pharmacokinetics in stable renal transplant patients. , 2009, Transplantation proceedings.

[4]  R. Takahashi.  Edaravone in ALS , 2009, Experimental Neurology.

[5]  S. Ohno,et al.  Determination of mRNA Expression of Human UDP-Glucuronosyltransferases and Application for Localization in Various Human Tissues by Real-Time Reverse Transcriptase-Polymerase Chain Reaction , 2009, Drug Metabolism and Disposition.

[6]  Aleksandra Galetin,et al.  Prediction of Drug Clearance by Glucuronidation from in Vitro Data: Use of Combined Cytochrome P450 and UDP-Glucuronosyltransferase Cofactors in Alamethicin-Activated Human Liver Microsomes , 2009, Drug Metabolism and Disposition.

[7]  M. Nakajima,et al.  Species Differences in UDP-Glucuronosyltransferase Activities in Mice and Rats , 2008, Drug Metabolism and Disposition.

[8]  Andrew Rowland,et al.  The “Albumin Effect” and Drug Glucuronidation: Bovine Serum Albumin and Fatty Acid-Free Human Serum Albumin Enhance the Glucuronidation of UDP-Glucuronosyltransferase (UGT) 1A9 Substrates but Not UGT1A1 and UGT1A6 Activities , 2008, Drug Metabolism and Disposition.

[9]  S. Todo,et al.  The novel antioxidant edaravone: from bench to bedside. , 2008, Cardiovascular therapeutics.

[10]  S. Nagar,et al.  Resveratrol (trans-Resveratrol, 3,5,4′-Trihydroxy-trans-stilbene) Glucuronidation Exhibits Atypical Enzyme Kinetics in Various Protein Sources , 2008, Drug Metabolism and Disposition.

[11]  A. Hishida.  Clinical analysis of 207 patients who developed renal disorders during or after treatment with edaravone reported during post-marketing surveillance , 2007, Clinical and Experimental Nephrology.

[12]  N. Perico,et al.  C-440T/T-331C polymorphisms in the UGT1A9 gene affect the pharmacokinetics of mycophenolic acid in kidney transplantation. , 2007, Pharmacogenomics.

[13]  Andrew Rowland,et al.  Binding of Inhibitory Fatty Acids Is Responsible for the Enhancement of UDP-Glucuronosyltransferase 2B7 Activity by Albumin: Implications for in Vitro-in Vivo Extrapolation , 2007, Journal of Pharmacology and Experimental Therapeutics.

[14]  S. Vermeire,et al.  The impact of uridine diphosphate–glucuronosyltransferase 1A9 (UGT1A9) gene promoter region single‐nucleotide polymorphisms T—275A and C—2152T on early mycophenolic acid dose‐interval exposure in de novo renal allograft recipients , 2005, Clinical pharmacology and therapeutics.

[15]  K. Knights,et al.  S-Naproxen and desmethylnaproxen glucuronidation by human liver microsomes and recombinant human UDP-glucuronosyltransferases (UGT): role of UGT2B7 in the elimination of naproxen. , 2005, British journal of clinical pharmacology.

[16]  Koujirou Yamamoto,et al.  Kidneys contribute to the extrahepatic clearance of propofol in humans, but not lungs and brain. , 2005, British journal of clinical pharmacology.

[17]  R. Laufer,et al.  DETERMINATION OF DRUG GLUCURONIDATION AND UDP-GLUCURONOSYLTRANSFERASE SELECTIVITY USING A 96-WELL RADIOMETRIC ASSAY , 2005, Drug Metabolism and Disposition.

[18]  A. Prémaud,et al.  IDENTIFICATION OF THE UDP-GLUCURONOSYLTRANSFERASE ISOFORMS INVOLVED IN MYCOPHENOLIC ACID PHASE II METABOLISM , 2005, Drug Metabolism and Disposition.

[19]  D. Greenblatt,et al.  Evaluation of 3'-azido-3'-deoxythymidine, morphine, and codeine as probe substrates for UDP-glucuronosyltransferase 2B7 (UGT2B7) in human liver microsomes: specificity and influence of the UGT2B7*2 polymorphism. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[20]  M. Nakajima,et al.  Glucuronidation of etoposide in human liver microsomes is specifically catalyzed by UDP-glucuronosyltransferase 1A1. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[21]  C. Klaassen,et al.  Tissue mRNA expression of the rat UDP-glucuronosyltransferase gene family. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[22]  D. Greenblatt,et al.  Serotonin (5-hydroxytryptamine) glucuronidation in vitro : assay development, human liver microsome activities and species differences , 2003, Xenobiotica; the fate of foreign compounds in biological systems.

[23]  J. Magdalou,et al.  The Human UDP-Glucuronosyltransferases: Structural Aspects and Drug Glucuronidation , 2003, Drug metabolism reviews.

[24]  T. Tracy,et al.  Atypical kinetic profiles in drug metabolism reactions. , 2002, Drug metabolism and disposition: the biological fate of chemicals.

[25]  J. Miners,et al.  The glucuronidation of mycophenolic acid by human liver, kidney and jejunum microsomes. , 2001, British journal of clinical pharmacology.

[26]  B. Burchell,et al.  Evidence for significant differences in microsomal drug glucuronidation by canine and human liver and kidney. , 2001, Drug metabolism and disposition: the biological fate of chemicals.

[27]  M. Murasaki,et al.  Phase I Clinical Study of MCI-186 (Edaravone, 3-methyl-1-phenyl-2-pyrazolin-5-one) in Healthy Volunteers , 1998 .

[28]  B. Burchell,et al.  Drug glucuronidation by human renal UDP-glucuronosyltransferases. , 1998, Biochemical pharmacology.

[29]  Y. Kumagai,et al.  A Pharmacokinetic Study of MCI-186, a Novel Drug for Cerebrovascular Disease in Elderly and Young Healthy Subjects , 1997 .

[30]  H. Nakai,et al.  Pharmacokinetic Studies of 3-Methyl-1-phenyl-2-pyrazolin-5-one-(MCI-186): Metabolism in Rats, Dogs and Human , 1996 .

[31]  T. Iyanagi,et al.  Drug-responsive and tissue-specific alternative expression of multiple first exons in rat UDP-glucuronosyltransferase family 1 (UGT1) gene complex. , 1995, Journal of biochemistry.

[32]  Yong Zhu,et al.  Synthesis of two metabolites of edaravone , 2010 .

[33]  K. Knights,et al.  Evidence that unsaturated fatty acids are potent inhibitors of renal UDP-glucuronosyltransferases (UGT): kinetic studies using human kidney cortical microsomes and recombinant UGT1A9 and UGT2B7. , 2004, Biochemical pharmacology.

[34]  R. Tukey,et al.  Human UDP-glucuronosyltransferases: metabolism, expression, and disease. , 2000, Annual review of pharmacology and toxicology.

[35]  M. Manns,et al.  UDP-glucuronosyltransferase activity in human liver and colon. , 1999, Gastroenterology.