Identification of Uridine 5′-Diphosphate-Glucuronosyltransferases Responsible for the Glucuronidation of Mirabegron, a Potent and Selective β3-Adrenoceptor Agonist, in Human Liver Microsomes

  • Kentaro KonishiEmail author
  • Daisuke Tenmizu
  • Shin Takusagawa
Original Research Article


Background and Objectives

Mirabegron is cleared by multiple mechanisms, including drug-metabolizing enzymes. One of the most important clearance pathways is direct glucuronidation. In humans, M11 (O-glucuronide), M13 (carbamoyl-glucuronide), and M14 (N-glucuronide) have been identified, of which M11 is one of the major metabolites in human plasma. The objective of this study was to identify the uridine 5′-diphosphate (UDP)-glucuronosyltransferase (UGT) isoform responsible for the direct glucuronidation of mirabegron using human liver microsomes (HLMs) and recombinant human UGTs (rhUGTs).


Reaction mixtures contained 1–1000 μM mirabegron, 8 mM MgCl2, alamethicin (25 μg/mL), 50 mM Tris–HCl buffer (pH 7.5), human liver microsome (HLM) or rhUGT (1.0 mg protein/mL), and 2 mM UDP-glucuronic acid in a total volume of 200 μL for 120 min at 37 °C. HLMs from 16 individuals were used for the correlation study, and mefenamic acid and propofol were used for the inhibition study.


Regarding M11 formation, rhUGT2B7 showed high activity among the rhUGTs tested (11.3 pmol/min/mg protein). This result was supported by the correlation between M11 formation activity and UGT2B7 marker enzyme activity (3-glucuronidation of morphine, r 2 = 0.330, p = 0.020) in individual HLMs; inhibition by mefenamic acid in pooled HLMs (IC50 = 22.8 μM); and relatively similar K m values between pooled HLMs and rhUGT2B7 (1260 vs. 486 μM). Regarding M13 and M14 formation, rhUGT1A3 and rhUGT1A8 showed high activity among the rhUGTs tested, respectively.


UGT2B7 is the main catalyst of M11 formation in HLMs. Regarding M13 and M14 formation, UGT1A3 and UGT1A8 are strong candidates for glucuronidation, respectively.



The authors would like to sincerely thank Mr. Tadashi Hashimoto, Dr. Kiyoshi Noguchi, and Dr. Takashi Usui for their useful suggestions about the experiments and Dr. Toshifumi Shiraga, Dr. Tsuyoshi Minematsu, Dr. Yasuhisa Nagasaka, Dr. Takafumi Iwatsubo, Dr. Yoichi Naritomi, Mr. Aiji Miyashita, and Dr. Kenji Tabata for their contribution to this study.

Author Contributions

Participated in research design Mr. Kentaro Konishi, Dr. Daisuke Tenmizu, and Dr. Shin Takusagawa. Conducted experiments Mr. Kentaro Konishi. Performed data analysis Mr. Kentaro Konishi, Dr. Daisuke Tenmizu, and Dr. Shin Takusagawa. Wrote or contributed to the writing of the manuscript Mr. Kentaro Konishi, Dr. Daisuke Tenmizu, and Dr. Shin Takusagawa

Compliance with Ethical Standards

Conflict of interest

Mr. Kentaro Konishi, Dr. Daisuke Tenmizu and  Dr. Shin Takusagawa are employees of Astellas Pharma, Japan.


This study was sponsored by Astellas Pharma, Japan. Editorial support was funded by Astellas Pharma, Japan.


  1. 1.
    Takasu T, Ukai M, Sato S, Matsui T, Nagase I, Maruyama T, et al. Effect of (R)-2-(2-aminothiazol-4-yl)-4′-{2-[(2-hydroxy-2-phenylethyl)amino]ethyl} acetanilide (YM178), a novel selective beta3-adrenoceptor agonist, on bladder function. J Pharmacol Exp Ther. 2007;321(2):642–7. Scholar
  2. 2.
    Yamaguchi O, Chapple CR. Beta3-adrenoceptors in urinary bladder. Neurourol Urodyn. 2007;26(6):752–6. Scholar
  3. 3.
    Herschorn S, Barkin J, Castro-Diaz D, Frankel JM, Espuna-Pons M, Gousse AE, et al. A phase III, randomized, double-blind, parallel-group, placebo-controlled, multicentre study to assess the efficacy and safety of the beta(3) adrenoceptor agonist, mirabegron, in patients with symptoms of overactive bladder. Urology. 2013;82(2):313–20. Scholar
  4. 4.
    Khullar V, Amarenco G, Angulo JC, Cambronero J, Hoye K, Milsom I, et al. Efficacy and tolerability of mirabegron, a beta(3)-adrenoceptor agonist, in patients with overactive bladder: results from a randomised European–Australian phase 3 trial. Eur Urol. 2013;63(2):283–95. Scholar
  5. 5.
    Nitti VW, Khullar V, van Kerrebroeck P, Herschorn S, Cambronero J, Angulo JC, et al. Mirabegron for the treatment of overactive bladder: a prespecified pooled efficacy analysis and pooled safety analysis of three randomised, double-blind, placebo-controlled, phase III studies. Int J Clin Pract. 2013;67(7):619–32. Scholar
  6. 6.
    Yamaguchi O, Marui E, Kakizaki H, Homma Y, Igawa Y, Takeda M, et al. Phase III, randomised, double-blind, placebo-controlled study of the beta3-adrenoceptor agonist mirabegron, 50 mg once daily, in Japanese patients with overactive bladder. BJU Int. 2014;113(6):951–60. Scholar
  7. 7.
    Takusagawa S, van Lier JJ, Suzuki K, Nagata M, Meijer J, Krauwinkel W, et al. Absorption, metabolism and excretion of [(14)C]mirabegron (YM178), a potent and selective beta(3)-adrenoceptor agonist, after oral administration to healthy male volunteers. Drug Metab Dispos. 2012;40(4):815–24. Scholar
  8. 8.
    Takusagawa S, Yajima K, Miyashita A, Uehara S, Iwatsubo T, Usui T. Identification of human cytochrome P450 isoforms and esterases involved in the metabolism of mirabegron, a potent and selective beta3-adrenoceptor agonist. Xenobiotica. 2012;42(10):957–67. Scholar
  9. 9.
    Krauwinkel W, van Dijk J, Schaddelee M, Eltink C, Meijer J, Strabach G, et al. Pharmacokinetic properties of mirabegron, a beta3-adrenoceptor agonist: results from two phase I, randomized, multiple-dose studies in healthy young and elderly men and women. Clin Ther. 2012;34(10):2144–60. Scholar
  10. 10.
    Eltink C, Lee J, Schaddelee M, Zhang W, Kerbusch V, Meijer J, et al. Single dose pharmacokinetics and absolute bioavailability of mirabegron, a beta(3)-adrenoceptor agonist for treatment of overactive bladder. Int J Clin Pharmacol Ther. 2012;50(11):838–50. Scholar
  11. 11.
    Fisher MB, Campanale K, Ackermann BL, VandenBranden M, Wrighton SA. In vitro glucuronidation using human liver microsomes and the pore-forming peptide alamethicin. Drug Metab Dispos. 2000;28(5):560–6.PubMedGoogle Scholar
  12. 12.
    Coffman BL, Rios GR, King CD, Tephly TR. Human UGT2B7 catalyzes morphine glucuronidation. Drug Metab Dispos. 1997;25(1):1–4.PubMedGoogle Scholar
  13. 13.
    Ohno S, Kawana K, Nakajin S. Contribution of UDP-glucuronosyltransferase 1A1 and 1A8 to morphine-6-glucuronidation and its kinetic properties. Drug Metab Dispos. 2008;36(4):688–94. Scholar
  14. 14.
    Joo J, Kim YW, Wu Z, Shin JH, Lee B, Shon JC, et al. Screening of non-steroidal anti-inflammatory drugs for inhibitory effects on the activities of six UDP-glucuronosyltransferases (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) using LC–MS/MS. Biopharm Drug Dispos. 2015;36(4):258–64. Scholar
  15. 15.
    Mano Y, Usui T, Kamimura H. Inhibitory potential of nonsteroidal anti-inflammatory drugs on UDP-glucuronosyltransferase 2B7 in human liver microsomes. Eur J Clin Pharmacol. 2007;63(2):211–6. Scholar
  16. 16.
    Mano Y, Usui T, Kamimura H. Predominant contribution of UDP-glucuronosyltransferase 2B7 in the glucuronidation of racemic flurbiprofen in the human liver. Drug Metab Dispos. 2007;35(7):1182–7. Scholar
  17. 17.
    Mano Y, Usui T, Kamimura H. The UDP-glucuronosyltransferase 2B7 isozyme is responsible for gemfibrozil glucuronidation in the human liver. Drug Metab Dispos. 2007;35(11):2040–4. Scholar
  18. 18.
    Mano Y, Usui T, Kamimura H. Contribution of UDP-glucuronosyltransferases 1A9 and 2B7 to the glucuronidation of indomethacin in the human liver. Eur J Clin Pharmacol. 2007;63(3):289–96. Scholar
  19. 19.
    Walsky RL, Bauman JN, Bourcier K, Giddens G, Lapham K, Negahban A, et al. Optimized assays for human UDP-glucuronosyltransferase (UGT) activities: altered alamethicin concentration and utility to screen for UGT inhibitors. Drug Metab Dispos. 2012;40(5):1051–65. Scholar
  20. 20.
    Zhang D, Chando TJ, Everett DW, Patten CJ, Dehal SS, Humphreys WG. In vitro inhibition of UDP glucuronosyltransferases by atazanavir and other HIV protease inhibitors and the relationship of this property to in vivo bilirubin glucuronidation. Drug Metab Dispos. 2005;33(11):1729–39. Scholar
  21. 21.
    Teijlingen R, Meijer J, Takusagawa S, Gelderen M, Beld C, Usui T. Development and validation of LC–MS/MS methods for the determination of mirabegron and its metabolites in human plasma and their application to a clinical pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci. 2012;887–888:102–11. Scholar
  22. 22.
    Houston JB, Kenworthy KE. In vitro-in vivo scaling of CYP kinetic data not consistent with the classical Michaelis–Menten model. Drug Metab Dispos. 2000;28(3):246–54.PubMedGoogle Scholar
  23. 23.
    Rowland A, Gaganis P, Elliot DJ, Mackenzie PI, Knights KM, Miners JO. Binding of inhibitory fatty acids is responsible for the enhancement of UDP-glucuronosyltransferase 2B7 activity by albumin: implications for in vitro-in vivo extrapolation. J Pharmacol Exp Ther. 2007;321(1):137–47. Scholar
  24. 24.
    Court MH, Zhang X, Ding X, Yee KK, Hesse LM, Finel M. Quantitative distribution of mRNAs encoding the 19 human UDP-glucuronosyltransferase enzymes in 26 adult and 3 fetal tissues. Xenobiotica. 2012;42(3):266–77. Scholar
  25. 25.
    Harbourt DE, Fallon JK, Ito S, Baba T, Ritter JK, Glish GL, et al. Quantification of human uridine-diphosphate glucuronosyl transferase 1A isoforms in liver, intestine, and kidney using nanobore liquid chromatography–tandem mass spectrometry. Anal Chem. 2012;84(1):98–105. Scholar
  26. 26.
    Bhasker CR, McKinnon W, Stone A, Lo AC, Kubota T, Ishizaki T, et al. Genetic polymorphism of UDP-glucuronosyltransferase 2B7 (UGT2B7) at amino acid 268: ethnic diversity of alleles and potential clinical significance. Pharmacogenetics. 2000;10(8):679–85.CrossRefPubMedGoogle Scholar
  27. 27.
    Kiang TK, Ensom MH, Chang TK. UDP-glucuronosyltransferases and clinical drug–drug interactions. Pharmacol Ther. 2005;106(1):97–132. Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Analysis and Pharmacokinetics Research Laboratories, Drug Discovery ResearchAstellas Pharma Inc.Tsukuba-shiJapan
  2. 2.Clinical Pharmacology, DevelopmentAstellas Pharma Inc.TokyoJapan

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