CYP21-catalyzed production of the long-term urinary metandienone metabolite 17β-hydroxymethyl-17α-methyl-18-norandrosta-1,4,13-trien-3-one: A contribution to the fight against doping

Biological Chemistry

Volumn 391, Issue 1, 2010, Pages 119-127

  Zöllner, Andy ^a   Parr, Maria Kristina ^b   Drǎgan, Cǎlin Aurel ^a   Dräs, Stefan ^a   Schlörer, Nils ^c   Peters, Frank T ^d,e   Maurer, Hans H ^d   Schänzer, Wilhelm ^b   Bureik, Matthias ^a  

^a PomBioTech GmbH   (Germany)

^b GERMAN SPORT UNIVERSITY COLOGNE   (Germany)

^c UNIVERSITY OF COLOGNE   (Germany)

^d SAARLAND UNIVERSITY   (Germany)

^e FRIEDRICH SCHILLER UNIVERSITY JENA   (Germany)

Author keywords

Anabolic steroids; CYP21; CYP3A4; Doping; Metabolite; Metandienone; Schizosaccharomyces pombe; Whole cell biotransformation.

Indexed keywords

17BETA HYDROXYMETHYL 17ALPHA METHYL 18 NORANDROSTA 1,4,13 TRIEN 3 ONE; ALDOSTERONE SYNTHASE; CYTOCHROME P450 11B1; CYTOCHROME P450 17; CYTOCHROME P450 2B6; CYTOCHROME P450 2C19; CYTOCHROME P450 2C9; CYTOCHROME P450 2D6; CYTOCHROME P450 3A4; CYTOCHROME P450 3A7; DRUG METABOLITE; METANDIENONE; STEROID; STEROID 21 MONOOXYGENASE; UNCLASSIFIED DRUG;

ARTICLE; BIOTRANSFORMATION; CONTROLLED STUDY; DOPING; DRUG ABUSE; FERMENTATION; FUNGAL STRAIN; HYDROXYLATION; NONHUMAN; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; PRIORITY JOURNAL; PURIFICATION; SCHIZOSACCHAROMYCES POMBE; STRUCTURE ANALYSIS; URINALYSIS; URINE;

ANABOLIC AGENTS; BIOTRANSFORMATION; CYTOCHROME P-450 CYP3A; DOPING IN SPORTS; HUMANS; MASS SPECTROMETRY; METHANDROSTENOLONE; NORSTEROIDS; NUCLEAR MAGNETIC RESONANCE, BIOMOLECULAR; RECOMBINANT PROTEINS; SCHIZOSACCHAROMYCES; STEROID 21-HYDROXYLASE; SUBSTANCE ABUSE DETECTION;

SCHIZOSACCHAROMYCES POMBE; SCHIZOSACCHAROMYCETACEAE;

EID: 75149179496     PISSN: 14316730     EISSN: 14374315     Source Type: Journal    
DOI: 10.1515/BC.2010.002     Document Type: Article

References (46)

1. Alfa, C., Fantes, P., Hyams, J., McLeod, M., and Warbrick, E. (1993). Experiments with Fission Yeast. A Laboratory Course Manual (Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press).
2. Bernhardt, R. (2005). Cytochromes P-450. In: Encyclopedia of Biological Chemistry, Vol.1, W. Lennarz, M. Lane, P. Modrich, J. Dixon, E. Carafoli, J. Exton, D. Cleveland, eds. (New York, USA: Academic Press), pp. 544-549.
3. Bernhardt, R. (2006). Cytochromes P450 as versatile biocatalysts. J. Biotechnol. 124, 128-145.
4. Bureik, M., Schiffler, B., Hiraoka, Y., Vogel, F., and Bernhardt, R. (2002). Functional expression of human mitochondrial CYP11B2 in fission yeast and identification of a new internal electron transfer protein, etp1. Biochemistry 41, 2311-2321.
5. Cardoso, C.C., Bornstein, S.R., and Hornsby, P.J. (2009). New methods for investigating experimental human adrenal tumorigenesis. Mol. Cell. Endocrinol. 300, 175-179.
6. Dragan, C.A., Zearo, S., Hannemann, F., Bernhardt, R., and Bureik, M. (2005). Efficient conversion of 11-deoxycortisol to cortisol (hydrocortisone) by recombinant fission yeast Schizosaccharomyces pombe. FEMS Yeast Res. 5, 621-625.
7. Dragan, C.A., Blank, L.M., and Bureik, M. (2006a). Increased TCA cycle activity and reduced oxygen consumption during cytochrome P450-dependent biotransformation in fission yeast. Yeast 23, 779-794.
8. Dragan, C.A., Hartmann, R.W., and Bureik, M. (2006b). A fission yeast-based test system for the determination of IC50 values of anti-prostate tumor drugs acting on CYP21. J. Enzym. Inhib. Med. Chem. 21, 547-556.
9. Forest, M.G. (2004). Recent advances in the diagnosis and management of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Hum. Reprod. Update 10, 469-485.
10. Guengerich, F.P. (2005). Human cytochrome P450 enzymes. In: Cytochrome P450 - Structure, Mechanism, and Biochemistry, P.R. Ortiz de Montellano, ed. (New York, USA: Kluwer Academic/ Plenum Publishers), pp. 377-530.
11. Hannemann, F., Bichet, A., Ewen, K.M., and Bernhardt, R. (2007). Cytochrome P450 systems - biological variations of electron transport chains. Biochim. Biophys. Acta 1770, 330-334.
12. Ingelman-Sundberg, M., Sim, S.C., Gomez, A., and Rodriguez-Antona, C. (2007). Influence of cytochrome P450 polymorphisms on drug therapies: pharmacogenetic, pharmacoepigenetic and clinical aspects. Pharmacol. Ther. 116, 496-526.
13. Isin, E.M. and Guengerich, F.P. (2007). Complex reactions catalyzed by cytochrome P450 enzymes. Biochim. Biophys. Acta 1770, 314-329.
14. Jansen, M.L.A., Krook, D.J.J., De Graaf, K., van Dijken, J.P., Pronk, J.T., and de Winde, J.H. (2006). Physiological characterization and fed-batch production of an extracellular maltase of Schizosaccharomyces pombe CBS 356. FEMS Yeast Res. 6, 888-901.
15. Kayes-Wandover, K.M. and White, P.C. (2000). Steroidogenic enzyme gene expression in the human heart. J. Clin. Endocrinol. Metab. 85, 2519-2525.
16. Korzekwa, K.R., Krishnamachary, N., Shou, M., Ogai, A., Parise, R.A., Rettie, A.E., Gonzalez, F.J., and Tracy, T.S. (1998). Evaluation of atypical cytochrome P450 kinetics with two-substrate models: evidence that multiple substrates can simultaneously bind to cytochrome P450 active sites. Biochemistry 37, 4137-4147.
17. Lin, Y., Lu, P., Tang, C., Mei, Q., Sandig, G., Rodrigues, A.D., Rushmore, T.H., and Shou, M. (2001). Substrate inhibition kinetics for cytochrome P450-catalyzed reactions. Drug Metab. Dispos. 29, 368-374. (Pubitemid 32275548)
18. Lorence, M.C., Trant, J.M., Mason, J.I., Bhasker, C.R., Fujii-Kuriyama, Y., Estabrook, R.W., and Waterman, M.R. (1989). Expression of a full-length cDNA encoding bovine adrenal cytochrome P450C21. Arch. Biochem. Biophys. 273, 79-88.
19. Losson, R. and Lacroute, F. (1983). Plasmids carrying the yeast OMP decarboxylase structural and regulatory genes: transcription regulation in a foreign environment. Cell 32, 371-377.
20. Martinez-Jimenez, C.P., Gomez-Lechon, M.J., Castell, J.V., and Jover, R. (2005). Transcriptional regulation of the human hepatic CYP3A4: identification of a new distal enhancer region responsive to CCAAT/enhancer-binding protein b isoforms (liver activating protein and liver inhibitory protein). Mol. Pharmacol. 67, 2088-2101.
21. Marynick, S.P., Chakmakjian, Z.H., McCaffree, D.L., and Herndon, J.H. (1983). Androgen excess in cystic acne. N. Engl. J. Med. 308, 981-986.
22. Maundrell, K. (1990). nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine. J. Biol. Chem. 265, 10857-10864.
23. Moreno, S., Klar, A., and Nurse, P. (1991). Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 194, 795-823.
24. Parr, M.K., Opfermann, G., Piper, T., Schlörer, N., Fussholler, G., Thomas, A., Thevis, M., and Schanzer, W. (2007). Characterization of Steroid Metabolites Recently Detected in Doping Control Assays, Vol.15 (Cologne, Germany: Sportverlag Strauss).
25. Peters, F.T., Dragan, C.A., Wilde, D.R., Meyer, M.R., Zapp, J., Bureik, M., and Maurer, H.H. (2007). Biotechnological synthesis of drug metabolites using human cytochrome P450 2D6 heterologously expressed in fission yeast exemplified for the designer drug metabolite 49-hydroxymethyl-a- pyrrolidinobutyrophenone. Biochem. Pharmacol. 74, 511-520.
26. Peters, F.T., Dragan, C.A., Kauffels, A., Schwaninger, A.E., Zapp, J., Bureik, M., and Maurer, H.H. (2009a). Biotechnological synthesis of the designer drug metabolite 49-hydroxymethyl-a-pyrrolidinohexanophenone in fission yeast heterologously expressing human cytochrome P450 2D6 - a versatile alternative to multistep chemical synthesis. J. Anal. Toxicol. 33, 190-197.
27. Peters, F.T., Dragan, C.A., Schwaninger, A.E., Sauer, C., Zapp, J., Bureik, M., and Maurer, H.H. (2009b). Use of fission yeast heterologously expressing human cytochrome P450 2B6 in biotechnological synthesis of the designer drug metabolite N-(1-phenylcyclohexyl)-2-hydroxyethanamine. Forensic Sci. Int. 184, 69-73.
28. Pirmohamed, M. and Park, B.K. (2003). Cytochrome P450 enzyme polymorphisms and adverse drug reactions. Toxicology 192, 23-32.
29. Rendic, S., Nolteernsting, E., and Schanzer, W. (1999). Metabolism of anabolic steroids by recombinant human cytochrome P450 enzymes. Gas chromatographic-mass spectrometric determination of metabolites. J. Chromatogr. B Biomed. Sci. Appl. 735, 73-83.
30. Reznik, Y., Dehennin, L., Coffin, C., Mahoudeau, J., and Leymarie, P. (2001). Urinary nandrolone metabolites of endogenous origin in man: a confirmation by output regulation under human chorionic gonadotropin stimulation. J. Clin. Endocrinol. Metab. 86, 146-150.
31. Rodriguez-Antona, C., Jande, M., Rane, A., and Ingelman-Sundberg, M. (2005). Identification and phenotype characterization of two CYP3A haplotypes causing different enzymatic capacity in fetal livers. Clin. Pharmacol. Ther. 77, 259-270.
32. Rosenwaks, Z., Lee, P.A., Jones, G.S., Migeon, C.J., and Wentz, A.C. (1979). Attenuated form of congenital virilizing adrenalhyperplasia. J. Clin. Endocrinol. Metab. 49, 335-339.
33. Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd Edition (Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press).
34. Schänzer, W. (1996). Metabolism of anabolic androgenic steroids. Clin. Chem. 42, 1001-1020.
35. Schänzer, W. and Donike, M. (1993). Metabolism of anabolic-steroids in man - synthesis and use of reference substances for identification of anabolic-steroid metabolites. Anal. Chim. Acta 275, 23-48.
36. Schänzer, W., Geyer, H., and Donike, M. (1991). Metabolism of metandienone in man: identification and synthesis of conjugated excreted urinary metabolites, determination of excretion rates and gas chromatographic-mass spectrometric identification of bis-hydroxylated metabolites. J. Steroid Biochem. Mol. Biol. 38, 441-464.
37. Schänzer, W., Geyer, H., Fussholler, G., Halatcheva, N., Kohler, M., Parr, M.K., Guddat, S., Thomas, A., and Thevis, M. (2006). Mass spectrometric identification and characterization of a new long-term metabolite of metandienone in human urine. Rapid Commun. Mass Spectrom. 20, 2252-2258.
38. Schulze, J.J., Lorentzon, M., Ohlsson, C., Lundmark, J., Roh, H.K., Rane, A., and Ekstrom, L. (2008). Genetic aspects of epitestosterone formation and androgen disposition: influence of polymorphisms in CYP17 and UGT2B enzymes. Pharmacogenet. Genomics 18, 477-485.
39. Tajima, T., Okada, T., Ma, X.M., Ramsey, W.J., Bornstein, S.R., and Aguilera, G. (1999). Restoration of adrenal steroidogenesis by adenovirus-mediated transfer of human cytochrome P450 21- hydroxylase into the adrenal gland of 21-hydroxylase-deficient mice. Gene Ther. 6, 1898-1903.
40. Temeck, J.W., Pang, S., Nelson, C., and New, M.I. (1987). Genetic defects of steroidogenesis in premature pubarche. J. Clin. Endocrinol. Metab. 64, 609-617.
41. van Herwaarden, A.E., Wagenaar, E., van der Kruijssen, C.M.M., van Waterschoot, R.A.B., Smit, J.W., Song, J.Y., van der Valk, M.A., van Tellingen, O., van der Hoorn, J.W.A., Rosing, H., et al. (2007). Knockout of cytochrome P450 3A yields new mouse models for understanding xenobiotic metabolism. J. Clin. Invest. 117, 3583-3592.
42. Walsky, R.L. and Obach, R.S. (2004). Validated assays for human cytochrome P450 activities. Drug Metab. Dispos. 32, 647-660.
43. White, P.C. and Speiser, P.W. (2000). Congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Endocr. Rev. 21, 245-291.
44. Wu, D.A., Hu, M.C., and Chung, B.C. (1991). Expression and functional study of wild-type and mutant human cytochrome P450c21 in Saccharomyces cerevisiae. DNA Cell Biol. 103, 201-209.
45. Zhou, Z.F., Agarwal, V.R., Dixit, N., White, P., and Speiser, P.W. (1997). Steroid 21-hydroxylase expression and activity in human lymphocytes. Mol. Cell. Endocrinol. 127, 11-18.
46. Zöllner, A., Dragan, C.A., Pistorius, D., Muller, R., Bode, H.B., Peters, F.T., Maurer, H.H., and Bureik, M. (2009). Human CYP4Z1 catalyzes the in-chain hydroxylation of lauric acid and myristic acid. Biol. Chem. 390, 313-317.