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Plasma pharmacokinetics and oral bioavailability of 3,4,5,6-tetrahydrouridine, a cytidine deaminase inhibitor, in mice

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Abstract

Cytidine analogues such as cytosine arabinoside, gemcitabine, decitabine, 5-azacytidine, 5-fluoro-2′-deoxycytidine and 5-chloro-2′-deoxycytidine undergo rapid catabolism by cytidine deaminase (CD). 3,4,5,6-tetrahydrouridine (THU) is a potent CD inhibitor that has been applied preclinically and clinically as a modulator of cytidine analogue metabolism. However, THU pharmacokinetics has not been fully characterized, which has impaired the optimal preclinical evaluation and clinical use of THU. Therefore, we characterized the THU pharmacokinetics and bioavailability in mice. Mice were dosed with THU iv (100 mg/kg) or po (30, 100, or 300 mg/kg). Plasma and urine THU concentrations were quantitated with a validated LC-MS/MS assay. Plasma pharmacokinetic parameters were calculated compartmentally and non-compartmentally. THU, at 100 mg/kg iv had a 73 min terminal half-life and produced plasma THU concentrations >1 μg/ml, the concentration shown to effectively block deamination, for 4 h. Clearance was 9.1 ml/min/kg, and the distribution volume was 0.95 l/kg. Renal excretion accounted for 36–55% of the THU dose. A three-compartment model fit the iv THU data best. THU, at 100 mg/kg po, produced a concentration versus time profile with a plateau of approximately 10 μg/ml from 0.5–3 h, followed by a decline with an 85 min half-life. The oral bioavailability of THU was approximately 20%. The 20% oral bioavailability of THU is sufficient to produce and sustain, for several hours, plasma concentrations that inhibit CD. This suggests the feasibility of using THU to decrease elimination and first-pass metabolism of cytidine analogues by CD. THU pharmacokinetics are now being evaluated in humans.

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References

  1. Akaike H (1979) A Bayesian extension of the minimal AIC procedures of autoregressive model fitting. Biometrika 66:237–242

    Article  Google Scholar 

  2. Beumer JH, Eiseman JL, Parise RA, Joseph E, Holleran JL, Covey JM, Egorin MJ (2006) Pharmacokinetics, metabolism, and oral bioavailability of the DNA methyltransferase inhibitor 5-fluoro-2′-deoxycytidine in mice. Clin Cancer Res 12:7483–7491

    Article  PubMed  CAS  Google Scholar 

  3. Beumer JH, Parise RA, Newman EM, Doroshow JH, Synold TW, Lenz H, Egorin MJ (2006) Concentrations of the DNA methyltransferase inhibitor 5-fluoro-2′-deoxycytidine (FdCyd) & its metabolites 5-fluoro-2′-deoxyuridine (FdUrd), 5-fluorouracil (FU), 5-fluorouridine (FUrd), & 5-fluorocytosine (FC) in plasma of patients treated with FdCyd. Proc Am Soc Clin Oncol 25

  4. Camiener GW, Smith CG (1965) Studies of the enzymatic deamination of cytosine arabinoside. I. Enzyme distribution and species specificity. Biochem Pharmacol 14:1405–1416

    Article  PubMed  CAS  Google Scholar 

  5. Chou TC, Arlin Z, Clarkson BD, Phillips FS (1977) Metabolism of 1-beta-D-arabinofuranosylcytosine in human leukemic cells. Cancer Res 37:3561–3570

    PubMed  CAS  Google Scholar 

  6. Cohen RM, Wolfenden R (1971) Cytidine deaminase from Escherichia coli. Purification, properties and inhibition by the potential transition state analog 3,4,5,6-tetrahydrouridine. J Biol Chem 246:7561–7565

    PubMed  CAS  Google Scholar 

  7. ADAPT II user’s guide (1997) Pharmacokinetic/pharmacodynamic systems analysis software. University of Southern California, Los Angeles

  8. Dareer SM, Mulligan LT Jr., White V, Tillery K, Mellett LB, Hill DL (1977) Distribution of [3H]cytosine arabinoside and its products in mice, dogs, and monkeys and effect of tetrahydrouridine. Cancer Treat Rep 61:395–407

    PubMed  CAS  Google Scholar 

  9. BRENDA: The comprehensive enzyme information system . http://www brenda uni-koeln de/index.php4 (2007). Available at http://www.brenda.uni-koeln.de/index.php4 (1/Feb./2007)

  10. el Dareer SM, White V, Chen FP, Mellett LB, Hill DL (1976) Distribution of tetrahydrouridine in experimental animals. Cancer Treat Rep 60:1627–1631

    PubMed  CAS  Google Scholar 

  11. Eliopoulos N, Cournoyer D, Momparler RL (1998) Drug resistance to 5-aza-2′-deoxycytidine, 2′,2′-difluorodeoxycytidine, and cytosine arabinoside conferred by retroviral-mediated transfer of human cytidine deaminase cDNA into murine cells. Cancer Chemother Pharmacol 42:373–378

    Article  PubMed  CAS  Google Scholar 

  12. Fenaux P (2005) Inhibitors of DNA methylation: beyond myelodysplastic syndromes. Nat Clin Pract Oncol 2(Suppl 1):S36-S44

    Article  PubMed  CAS  Google Scholar 

  13. Freireich EJ, Gehan EA, Rall DP, Schmidt LH, Skipper HE (1966) Quantitative comparison of toxicity of anticancer agents in mouse, rat, hamster, dog, monkey, and man. Cancer Chemother Rep 50:219–244

    PubMed  CAS  Google Scholar 

  14. Furner RL, Mellett LB (1975) Inhibition of deamination of 14C-cytosine arabinoside (NSC-63878): a useful biologic assay for tetrahydrouridine (NSC-112907). Cancer Chemother Rep 59:717–720

    PubMed  CAS  Google Scholar 

  15. Furner RL, Mellett LB, Herren TC (1975) Influence of tetrahydrouridine on the phosphorylation of 1-beta-D-arabinofuranosyl-cytosine (ara-C) by enzymes from solid tumors in vitro. J Pharmacol Exp Ther 194:103–110

    PubMed  CAS  Google Scholar 

  16. Goldenthal EI, Cookson KM, Geil RG, Wazeter FX (1974) Preclinical toxicologic evaluation of tetrahydrouridine (NSC-112907) in beagle dogs and rhesus monkeys. Cancer Chemother Rep 3(5):15–16

    Google Scholar 

  17. Grant S, Bhalla K, McCrady C (1991) Effect of tetrahydrouridine and deoxytetrahydrouridine on the interaction between 2′-deoxycytidine and 1-beta-D-arabinofuranosylcytosine in human leukemia cells. Leuk Res 15:205–213

    Article  PubMed  CAS  Google Scholar 

  18. Greer S, Schwade J, Marion HS (1995) Five-chlorodeoxycytidine and biomodulators of its metabolism result in fifty to eighty percent cures of advanced EMT-6 tumors when used with fractionated radiation. Int J Radiat Oncol Biol Phys 32:1059–1069

    PubMed  CAS  Google Scholar 

  19. Hale JT, Bigelow JC, Mathews LA, McCormack JJ (2002) Analytical and pharmacokinetic studies with 5-chloro-2′-deoxycytidine. Biochem Pharmacol 64:1493–1502

    Article  PubMed  CAS  Google Scholar 

  20. Ho DH (1973) Distribution of kinase and deaminase of 1-beta-D-arabinofuranosylcytosine in tissues of man and mouse. Cancer Res 33:2816–2820

    PubMed  CAS  Google Scholar 

  21. Ho DH, Bodey GP, Hall SW, Benjamin RS, Brown NS, Freireich EJ, Loo TL (1978) Clinical pharmacology of tetrahydrouridine. J Clin Pharmacol 18:259–265

    PubMed  CAS  Google Scholar 

  22. Ho DH, Carter CJ, Brown NS, Hester J, McCredie K, Benjamin RS, Freireich EJ, Bodey GP (1980) Effects of tetrahydrouridine on the uptake and metabolism of 1-beta-D-arabinofuranosylcytosine in human normal and leukemic cells. Cancer Res 40:2441–2446

    CAS  Google Scholar 

  23. Kaye SB (1998) New antimetabolites in cancer chemotherapy and their clinical impact. Br J Cancer 78(Suppl 3):1–7

    PubMed  CAS  Google Scholar 

  24. Klubes P, Geffen DB, Cysyk RL (1986) Comparison of the bioavailability of uridine in mice after either oral or parenteral administration. Cancer Chemother Pharmacol 17:236–240

    Article  PubMed  CAS  Google Scholar 

  25. Kreis W, Chan K, Budman DR, Schulman P, Allen S, Weiselberg L, Lichtman S, Henderson V, Freeman J, Deere M (1988) Effect of tetrahydrouridine on the clinical pharmacology of 1-beta-D-arabinofuranosylcytosine when both drugs are coinfused over three hours. Cancer Res 48:1337–1342

    PubMed  CAS  Google Scholar 

  26. Kreis W, Woodcock TM, Gordon CS, Krakoff IH (1977) Tetrahydrouridine: physiologic disposition and effect upon deamination of cytosine arabinoside in man. Cancer Treat Rep 61:1347–1353

    PubMed  CAS  Google Scholar 

  27. Laliberte J, Marquez VE, Momparler RL (1992) Potent inhibitors for the deamination of cytosine arabinoside and 5-aza-2′-deoxycytidine by human cytidine deaminase. Cancer Chemother Pharmacol 30:7–11

    Article  PubMed  CAS  Google Scholar 

  28. Li Y, Looney GA, Kimler BF, Hurwitz A (1997) Opiate effects on 5-fluorouracil disposition in mice. Cancer Chemother Pharmacol 39:273–277

    Article  PubMed  CAS  Google Scholar 

  29. Lopez JA, Agarwal RP (1984) Acute cerebellar toxicity after high-dose cytarabine associated with CNS accumulation of its metabolite, uracil arabinoside. Cancer Treat Rep 68:1309–1310

    PubMed  CAS  Google Scholar 

  30. Marsh JH, Kreis W, Barile B, Akerman S, Schulman P, Allen SL, DeMarco LC, Schuster MW, Budman DR (1993) Therapy of refractory/relapsed acute myeloid leukemia and blast crisis of chronic myeloid leukemia with the combination of cytosine arabinoside, tetrahydrouridine, and carboplatin. Cancer Chemother Pharmacol 31:481–484

    Article  PubMed  CAS  Google Scholar 

  31. Mekras JA, Boothman DA, Greer SB (1985) Use of 5-trifluoromethyldeoxycytidine and tetrahydrouridine to circumvent catabolism and exploit high levels of cytidine deaminase in tumors to achieve DNA- and target-directed therapies. Cancer Res 45:5270–5280

    PubMed  CAS  Google Scholar 

  32. Mulligan LT Jr. (1970) A comparative study of the metabolism of 1-beta-D-arabinofuranosyl cytosine in various animal species. University of Alabama in Birmingham (thesis)

  33. Nand S, Messmore HL Jr., Patel R, Fisher SG, Fisher RI (1986) Neurotoxicity associated with systemic high-dose cytosine arabinoside. J Clin Oncol 4:571–575

    PubMed  CAS  Google Scholar 

  34. Neil GL, Moxley TE, Kuentzel SL, Manak RC, Hanka LJ (1975) Enhancement by tetrahydrouridine (NSC-112907) of the oral activity of 5-azacytidine (NSC-102816) in L1210 leukemic mice. Cancer Chemother Rep 59:459–465

    PubMed  Google Scholar 

  35. Neil GL, Moxley TE, Manak RC (1970) Enhancement by tetrahydrouridine of 1-beta-D-arabinofuranosylcytosine (cytarabine) oral activity in L1210 leukemic mice. Cancer Res 30:2166–2172

    PubMed  CAS  Google Scholar 

  36. Newman EM, Longmate J, Lenz H, Carroll M, Stalter S, Lim D, Raschko D, Shibata S, Somlo G, Doroshow J (2002) Phase I and clinical pharmacokinetic evaluation of the DNA methyltransferase inhibitor 5-fluoro-2′-deoxycytidine: a California Cancer Consortium study. Proc Am Soc Clin Oncol 21:108a

    Google Scholar 

  37. North TW, Mathews CK (1981) Tetrahydrouridine specifically facilitates deoxycytidine incorporation into herpes simplex virus DNA. J Virol 37:987–993

    PubMed  CAS  Google Scholar 

  38. Parise RA, Egorin MJ, Eiseman JL, Joseph E, Covey JM, Beumer JH (2007) Quantitative determination of the cytidine deaminase inhibitor tetrahydrouridine (THU) in mouse plasma by liquid chromatography/electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom 21:1991–1997

    Article  PubMed  CAS  Google Scholar 

  39. Peters GJ, van Groeningen CJ, Laurensse EJ, Lankelma J, Leyva A, Pinedo HM (1987) Uridine-induced hypothermia in mice and rats in relation to plasma and tissue levels of uridine and its metabolites. Cancer Chemother Pharmacol 20:101–108

    Article  PubMed  CAS  Google Scholar 

  40. Riva C, Barra Y, Carcassonne Y, Cano JP, Rustum Y (1992) Effect of tetrahydrouridine on metabolism and transport of 1-beta-D-arabinofuranosylcytosine in human cells. Chemotherapy 38:358–366

    Article  PubMed  CAS  Google Scholar 

  41. Shipley LA, Brown TJ, Cornpropst JD, Hamilton M, Daniels WD, Culp HW (1992) Metabolism and disposition of gemcitabine, and oncolytic deoxycytidine analog, in mice, rats, and dogs. Drug Metab Dispos 20:849–855

    PubMed  CAS  Google Scholar 

  42. Stoller RG, Myers CE, Chabner BA (1978) Analysis of cytidine deaminase and tetrahydrouridine interaction by use of ligand techniques. Biochem Pharmacol 27:53–59

    Article  PubMed  CAS  Google Scholar 

  43. Guidance for industry—bioanalytical method validation. http://www fda gov/cder/guidance/index htm (May/2001). Available at http://www.fda.gov/cder/guidance/index.htm (1/Feb./2007)

  44. Wong PP, Currie VE, Mackey RW, Krakoff IH, Tan CT, Burchenal JH, Young CW (1979) Phase I evaluation of tetrahydrouridine combined with cytosine arabinoside. Cancer Treat Rep 63:1245–1249

    PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Dr. B. Rao Vishnuvajjala for providing the D 4-THU internal standard, Diane Mazzei and her colleagues at the University of Pittsburgh Animal Facility for their expert assistance, and the University of Pittsburgh Cancer Institute Hematology/Oncology Writing Group for constructive suggestions regarding the manuscript.

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Authors and Affiliations

  1. Molecular Therapeutics/Drug Discovery Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, 15213, USA

    Jan H. Beumer, Julie L. Eiseman, Robert A. Parise, Erin Joseph, Robert S. Parker & Merrill J. Egorin

  2. Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Jan H. Beumer

  3. Department of Pharmacology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Julie L. Eiseman & Merrill J. Egorin

  4. Department of Chemical and Petroleum Engineering, School of Engineering, University of Pittsburgh, Pittsburgh, PA, 15218, USA

    Jeffry A. Florian Jr. & Robert S. Parker

  5. Department of Biomedical Engineering, University of Southern California, Los Angeles, CA, 90089, USA

    David Z. D’Argenio & Brittany Kay

  6. Toxicology and Pharmacology Branch, Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Rockville, MD, 20852, USA

    Joseph M. Covey

  7. Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Merrill J. Egorin

  8. Hillman Research Pavilion, University of Pittsburgh Cancer Institute, Room G27D, 5117 Centre Avenue, Pittsburgh, PA, 15213-1863, USA

    Jan H. Beumer

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  1. Jan H. Beumer
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Correspondence to Jan H. Beumer.

Additional information

Grant support: This work was supported by contract NO1-CM-52202 and grant P30-CA47904 from the National Cancer Institute; and grant P41-EB001978 awarded by the National Institute of Biomedical Imaging and Bioengineering.

WinNonlin software was provided as part of the Pharsight Academic Licensing Program.

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Beumer, J.H., Eiseman, J.L., Parise, R.A. et al. Plasma pharmacokinetics and oral bioavailability of 3,4,5,6-tetrahydrouridine, a cytidine deaminase inhibitor, in mice. Cancer Chemother Pharmacol 62, 457–464 (2008). https://doi.org/10.1007/s00280-007-0625-2

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