Skip to main content
SpringerLink
Log in

Investigations on the in vitro metabolism of five synthetic 19-norprogestins using hepatocyte suspensions isolated from five laboratory animal species

  • Published:
European Journal of Drug Metabolism and Pharmacokinetics Aims and scope Submit manuscript

Summary

Five synthetic progestins of the 19-nortestosterone type (norethisterone, NET; levonorgestrel, LN; gestodene, GEST; NET-3-oxime, NETO; norgestimate, NGM) were investigated in the in vitro hepatocyte model. Radiolabelled progestins were added to hepatocyte suspensions (3 × 106 cells/ml) freshly prepared from female rat, guinea pig, rabbit, dog (beagle) and cynomolgus monkey. Drug level decreases (NET, LN, GEST) and prodrug conversions (NETO, NGM) were followed by radiochromatography (HPLC) for 60 min. Li the case of NET and NETO the conversion into ethinyl estradiol (EE2) was quantified by RIA after HPLC separation. Half-lives of drug level decreases (t1/2), areas under the curves (AUC) and metabolic clearance rates (MCR) were estimated for all progestins.

For NETO and NGM the percentages of conversion into NET and LN were calculated, respectively, and levels of EE2 determined in the case of NET and NETO.

Rat hepatocytes showed an extremely high metabolic activity towards NET, LN and GEST resulting in tin values of below 2 min. Respective values for rabbit hepatocytes ranged from 5–8 min, whereas half-lives calculated for liver cells from guinea pig, dog and monkey were generally above 30 min. A drastic increase in t1/2 was found for NETO (as compared to NET) in hepatocytes from rat, rabbit and monkey but not from guinea pig. Dog hepatocytes degraded NETO about 3 times more rapidly than NET. NGM was degraded much fester than LN in hepatocytes from all species except the rat Liver cells from guinea pig and dog seem to be able to metabolize the 3-oxime group much more rapidly than hepatocytes from the other animal species. The lowest degree of prodrug conversion of 4% was observed for NGM and dog hepatocytes. Elevated EE2 levels were found in all experiments with NET and NETO. Results of NET, LN and GEST were compared with published in vivo experiments. No correlations were found for t1/2, MCR, and AUC.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Skanberg I. (1980): Metabolism of beta-2 adrenoceptor antagonists alpienolol and metoprolol in different species: in vitro and in vivo correlations. Acta. Pharm. Suecica, 17, 295–296.

    CAS  Google Scholar 

  2. Mizuma T., Machida M., Hayashi M., Awazu S. (1982): Correlation of drug conjugative metabolism rates between in vivo and in vitro glucuronidation and sulfation of p-nitrophenol as a model compound in rat. J. Pharmacobiol. Dyn., 5, 811–817.

    CAS  Google Scholar 

  3. Lin J.H., Hayashi M., Awazu S., Hanano M. (1978): Correlation between in vitro and in vivo drug metabolism rate: oxidation of ethoxybenzamide in rat. J. Pharmacokinet Biopharm., 6, 327–337.

    Article  CAS  PubMed  Google Scholar 

  4. Hümpel M., Sostarek D., Gieschen H., Labitzky Chr. (1989): Studies on the biotransformation of lonazolac, bromerguride, lisuride and terguride in laboratory animals and their hepatocytes. Xenobiotica, 19, 361–377.

    Article  PubMed  Google Scholar 

  5. Hümpel M., Düsterberg B., Beier S., Schuppler J., Günzel P., Elger W. (1986): The role of pharmacokinetics in preclinical safety studies of synthetic sex steroids. In: Gregoire A.T., Blye R.P. eds. Contraceptive Steroids, pp. 47–65.

  6. Glocklin V.C. (1981): General considerations for studies of the metabolism of drugs and other chemicals. 65th Annual FASEB Meeting, Atlanta, April 16.

  7. Calabrese E.J. (1984): Suitability of animal models for predictive toxicology: Theoretical and practical considerations. Drug Metab. Rev., 15, 505–523.

    Article  CAS  PubMed  Google Scholar 

  8. Meijer K.F., Keulemans K., Mulder G.J. (1981): Isolated perfused rat liver technique. Meth. Enzymol., 77, 81–93.

    Article  CAS  PubMed  Google Scholar 

  9. Moldéus P., Högberg J., Orrhenius S. (1978): Isolation and use of liver cells. Meth. Enzymol., 52, 60–71.

    Article  PubMed  Google Scholar 

  10. Guillouzo A., Begue J.P., Campion J.P., Gascoin M.-N., Guguen-Guillouzo C. (1985): Human hepatocyte cultures: a model of pbarmaco-toxicological studies. Xenobiotica, 15, 635–641.

    Article  CAS  PubMed  Google Scholar 

  11. Yamamoto T., Kitawaki J., Shiroshita K. et al. (1988): The confirmation of norethisterone aromatization in primary human hepatocytes by the vitafiber-II cell culture system. Acta. Obst. Gynaec. Jpn., 40, 87–89.

    CAS  Google Scholar 

  12. Peter H., Jung R., Bolt H.M., Oesch F. (1981): Norethisterone-4β, 5-oxide and levonorgestrel-4β, 5-oxide: formation in rat liver microsomal incubations and interference with microsomal epoxide hydrolase and cytoplasmic glutathione S-transferase. J. Steroid Biochem., 14, 83–90.

    Article  CAS  PubMed  Google Scholar 

  13. Düsterberg B., Hümpel M., Speck U. (1981): Terminal half-lives in plasma and bioavailability of norethisterone, levonorgestrel, cyproterone acetate and gestodene in rats, beagles and rhesus monkeys, Contraception, 24, 673–683.

    Article  PubMed  Google Scholar 

  14. Hümpel M. (1989): Comparative pharmacokinetics of selected contraceptive steroids in animal species and man In: Michal F. (ed) Safety Requirements for Contraceptive Steroids; Cambridge, Cambridge University Press, pp. 193–210.

    Google Scholar 

  15. Reese J.A., Byard J.L. (1981): Isolation and culture of adult hepatocytes from liver biopsies. In Vitro, 17, 935–940.

    Article  CAS  PubMed  Google Scholar 

  16. Hümpel M., Nieuweboer B., Wendt H., Speck U. (1979): Investigations of pharmacokinetics of ethinyl estradiol to specific consideration of a possible first-pass effect. Contraception, 19, 421–432.

    Article  PubMed  Google Scholar 

  17. Rodbard D. (1978): Statistical estimation of the minimal detectable concentration (“sensitivity”) for radioligand assays. Anal. Biochem., 90, 1–12.

    Article  CAS  PubMed  Google Scholar 

  18. Qi-Gui L. (1989): Investigations on the metabolism of five progestogens of the 19-nortestosterone type in various laboratory animal species. Dissertation at the Faculty of Pharmacy, Free University of Berlin.

  19. Quon C.Y., Stampfli H.F. (1985): Biochemical properties of blood esmolol esterase. Drug Metab. Dispos., 13, 420–424.

    CAS  PubMed  Google Scholar 

  20. Lee R.M., Livett B.H. (1967): A method for the colorimetric estimation of local anaesthetics containing ester link, and its use in the determination of esterase activity. Biochem. Pharmacol., 16, 1757–1765.

    Article  CAS  PubMed  Google Scholar 

  21. Günzel P., Putz B., Lehmann M., Hasan S.H., Hümpel M., El Etreby F. (1989): Steroid toxicology and the “pill”: comparative aspects of experimental test systems and the human. In: Dayan A.D.)., Paine A.J. (eds). Advances in Applied Toxicology. London, Tailor and Francis, pp. 19–49.

    Google Scholar 

  22. Qi-Gui L., Dunzhou L. (1987): Analysis of anti-implantational action of norethisterone-3-oxime. Contraception, 36, 667–676.

    Article  Google Scholar 

  23. Lin P., Yidi A., (1986): Morphological studies on the mechanism of antiimplantation and early pregnancy termination effect by norethisterone oxime in rats. Shengzhi Yo Biyun, 6, 38–41.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacology, Walter Reed Army Institute of Research, Washington, USA

    L. Qi-Gui

  2. Famify Planning Research Institute, Tonji Medical University, Wuhan, People’s Republic of China

    Z. Ming-Da

  3. Schering AG, Muellestr. 170-178, D-1000, Berlin, Bergkamen, Germany

    M. Hümpel

Authors
  1. L. Qi-Gui
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Z. Ming-Da
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Hümpel
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Qi-Gui, L., Ming-Da, Z. & Hümpel, M. Investigations on the in vitro metabolism of five synthetic 19-norprogestins using hepatocyte suspensions isolated from five laboratory animal species. European Journal of Drug Metabolism and Pharmacokinetics 16, 93–102 (1991). https://doi.org/10.1007/BF03189944

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03189944

Keywords

Search

Navigation