Development and validation of a high-throughput radiometric CYP3A4/5 inhibition assay using tritiated testosterone

Drug Metabolism and Disposition

Volumn 33, Issue 3, 2005, Pages 349-358

  Di Marco, Annalise ^a   Marcucci, Isabella ^a   Verdirame, Maria ^a   Pérez, José ^b   Sanchez, Manuel ^b   Peláez, Fernando ^b   Chaudhary, Ashok ^c   Laufer, Ralph ^a,d  

^a Merck Research Laboratories   (Italy)

^b Medical affairs Merck sharp and Dohme ltd   (Spain)

^c MERCK RESEARCH LABORATORIES   (United States)

^d IRBM   (Italy)

Author keywords

[No Author keywords available]

Indexed keywords

5BETA,6BETA EPOXY 17BETA HYDROXYANDROSTAN 3 ONE; 6BETA 3,3 ETHYLENEDIOXYANDROSTANE 5ALPHA,17BETA DIOL; 6BETA TESTOSTERONE; BROMOCRIPTINE; CYTOCHROME P450 3A4; CYTOCHROME P450 3A5; CYTOCHROME P450 INHIBITOR; KETOCONAZOLE; MICONAZOLE; MONOCLONAL ANTIBODY; NICARDIPINE; NIFEDIPINE; REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE; TESTOSTERONE; TRITIUM; UNCLASSIFIED DRUG;

ARTICLE; BETA CHAIN; CONTROLLED STUDY; DRUG POTENCY; ENZYME INHIBITION; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; HIGH THROUGHPUT SCREENING; HUMAN; HUMAN CELL; HYDROXYLATION; IC 50; INCUBATION TIME; INSTRUMENTATION; KINETICS; LABORATORY AUTOMATION; MASS SPECTROMETRY; PRIORITY JOURNAL; RADIOACTIVITY; RADIOMETRY; SENSITIVITY ANALYSIS; SEPARATION TECHNIQUE; SOLID PHASE EXTRACTION; STATISTICAL SIGNIFICANCE; TANDEM MASS SPECTROMETRY; TESTOSTERONE SYNTHESIS; TITRIMETRY; VALIDATION PROCESS;

ANTIBODIES, MONOCLONAL; AUTOMATION; CYTOCHROME P-450 ENZYME SYSTEM; ENZYME INHIBITORS; HUMANS; HYDROXYLATION; HYDROXYTESTOSTERONES; MICROSOMES, LIVER; RADIOMETRY; REPRODUCIBILITY OF RESULTS; TESTOSTERONE; TRITIUM; WATER;

EID: 14044275075     PISSN: 00909556     EISSN: None     Source Type: Journal    
DOI: 10.1124/dmd.104.002873     Document Type: Article

References (48)

1. Bertz RJ and Granneman GR (1997) Use of in vitro and in vivo data to estimate the likelihood of metabolic pharmacokinetic interactions. Clin Pharmacokinet 32:210-258.
2. Bjorkhem I (1972) On the rate-limiting step in microsomal hydroxylation of steroids. Eur J Biochem 27:354-363.
3. Bourrie M, Meunier V, Berger Y, and Fabre G (1996) Cytochrome P450 isoform inhibitors as a tool for the investigation of metabolic reactions catalyzed by human liver microsomes. J Pharmacol Exp Ther 277:321-332.
4. Campbell JA, Babcock JC, and Hogg JA (1958) 6-Methyl steroids in the androstane series. J Am Chem Soc 80:4717-4721.
5. Chauret N, Gauthier A, and Nicoll-Griffith DA (1998) Effect of common organic solvents on in vitro cytochrome P450-mediated metabolic activities in human liver microsomes. Drug Metab Dispos 26:1-4.
6. Chauret N, Tremblay N, Lackman RL, Gauthier JY, Silva JM, Marois J, Yergey JA, and Nicoll-Griffith DA (1999) Description of a 96-well plate assay to measure cytochrome P4503A inhibition in human liver microsomes using a selective fluorescent probe. Anal Biochem 276:215-226.
7. Cohen LH, Remley MJ, Raunig D, and Vaz AD (2003) In vitro drug interactions of cytochrome P450: an evaluation of fluorogenic to conventional substrates. Drug Metab Dispos 31:1005-1015.
8. Crespi CL and Stresser DM (2000) Fluorometric screening for metabolism-based drug-drug interactions. J Pharmacol Toxicol Methods 44:325-331.
9. Daly JW (1970) A simple radiometric assay for microsomal aryl hydroxylase activity. Anal Biochem 33:286-296.
10. De Lean A and Rodbard D (1979) Kinetics of cooperative binding, in Receptors: Comprehensive Treatise (O'Brien RD ed) vol. 1, pp 143-192, Plenum Publishing Corporation, New York.
11. Di Marco A, Marcucci I, Chaudhary A, Taliani M, and Laufer R (2005) Development and validation of a high-throughput radiometric CYP2C9 inhibition assay using tritiated diclofenac. Drug Metab Dispos 33:359-364.
12. Draper AJ, Madan A, Smith K, and Parkinson A (1998) Development of a non-high pressure liquid chromatography assay to determine testosterone hydroxylase (CYP3A) activity in human liver microsomes. Drug Metab Dispos 26:299-304.
13. Eagling VA, Tjia JF, and Back DJ (1998) Differential selectivity of cytochrorne P450 inhibitors against probe substrates inhuman and rat liver microsomes. Br J Clin Pharmacol 45:107-114.
14. Ekins S, Stresser DM, and Williams JA (2003) In vitro and pharmacophore insights into CYP3A enzymes. Trends Pharmacol Sci 24:161-166.
15. Gelb MH, Heimbrook DC, Malkonen P, and Sligar SG (1982) Stereochemistry and deuterium isotope effects in camphor hydroxylation by the cytochrome P450cam monooxygenase system. Biochemistry 21:370-377.
16. Guengerich FP (1999) Cytochrome P-450 3A4: regulation and role in drug metabolism. Annu Rev Pharmacol Toxicol 39:1-17.
17. Hayakawa T and Udenfriend S (1973) A simple radioisotope assay for microsomal aryl hydroxylase. Anal Biochem 51:501-509.
18. Hickman D, Wang JP, Wang Y, and Unadkat JD (1998) Evaluation of the selectivity of in vitro probes and suitability of organic solvents for the measurement of human cytochrome P450 monooxygenase activities. Drug Metab Dispos 26:207-215.
19. Ioannides C, ed. (1996) Cytochromes P450. Metabolic and Toxicological Aspects. CRC Press, Boca Raton, FL.
20. Kenworthy KE, Bloomer JC, Clarke SE, and Houston JB (1999) CYP3A4/5 drug interactions: correlation of 10 in vitro probe substrates. Br J Clin Pharmacol 48:716-727.
21. Kenworthy KE, Clarke SE, Andrews J, and Houston JB (2001) Multisite kinetic models for CYP3A4/5: simultaneous activation and inhibition of diazepam and testosterone metabolism. Drug Metab Dispos 29:1644-1651.
22. Krauser JA, Voehler M, Tseng LH, Schefer AB, Godejohann M, and Guengerich FP (2004) Testosterone 1 beta-hydroxylation by human cytochrome P450 3A4. Eur J Biochem 271:3962-3969.
23. Lin JH and Lu AY (1998) Inhibition and induction of cytochrome P450 and the clinical implications. Clin Pharmacokinet 35:361-390.
24. Liston AJ and Toft P (1969) Enol acetylation of alkylated D4-3-oxo steroids. A novel enonephenol transformation. J Org Chem 34:2288-2296.
25. Maenpaa J, Pelkonen O, Cresteil T, and Rane A (1993) The role of cytochrome P450 3A (CYP3A) isoform(s) in oxidative metabolism of testosterone and benzphetamine in human adult and fetal liver. J Steroid Biochem Mol Biol 44:61-67.
26. Mei Q, Tang C, Assang C, Lin Y, Slaughter D, Rodrigues AD, Baillie TA, Rushmore TH, and Shou M (1999) Role of a potent inhibitory monoclonal antibody to cytochrome P-450 3A4 in assessment of human drug metabolism. J Pharmacol Exp Ther 291:749-759.
27. Moody GC, Griffin SJ, Mather AN, McGinnity DF, and Riley RJ (1999) Fully automated analysis of activities catalysed by the major human liver cytochrome P450 (CYP) enzymes: assessment of human CYP inhibition potential. Xenobiotica 29:53-75.
28. Northrop DB (1982) Deuterium and tritium kinetic isotope effects on initial rates. Methods Enzymol 87:607-625.
29. Osawa Y and Coon MJ (1987) Radiometric assay for cytochrome P-450-catalyzed progesterone 16 alpha-hydroxylation and determination of an apparent isotope effect. Anal Biochem 164:355-361.
30. Reed KC and Ohno S (1976) Kinetic properties of human placental aromatase. Application of an assay measuring 3H2O release from 1beta,2beta-3H-androgens. J Biol Chem 251:1625-1631.
31. Riley RJ (2001) The potential pharmacological and lexicological impact of P450 screening. Curr Opin Drug Discovery Dev 4:45-54.
32. Rodbard D and Frazier GR (1975) Statistical analysis of radioligand assay data. Methods Enzymol 37:3-22.
33. Shou M, Lin Y, Lu P, Tang C, Mei Q, Cui D, Tang W, Ngui JS, Lin CC, Singh R, et al. (2001) Enzyme kinetics of cytochrome P450-mediated reactions. Curr Drug Metab 2:17-36.
34. Shou M, Lu T, Krausz KW, Sai Y, Yang T, Korzekwa KR, Gonzalez FJ, and Gelboin HV (2000) Use of inhibitory monoclonal antibodies to assess the contribution of cytochromes P450 to human drug metabolism. Eur J Pharmacol 394:199-209.
35. Stresser DM, Blanchard AP, Turner SD, Erve JC, Dandeneau AA, Miller VP, and Crespi CL (2000) Substrate-dependent modulation of CYP3A4/5 catalytic activity: analysis of 27 test compounds with four fluorometric substrates. Drug Metab Dispos 28:1440-1448.
36. Stresser DM, Turner SD, Blanchard AP, Miller VP, and Crespi CL (2002) Cytochrome P450 fluorometric substrates: identification of isoform-selective probes for rat CYP2D2 and human CYP3A4/5. Drug Metab Dispos 30:845-852.
37. Thompson EA Jr and Siiteri PK (1974) Utilization of oxygen and reduced nicotinamide adenine dinucleotide phosphate by human placental microsomes during aromatization of androstenedione. J Biol Chem 249:5364-5372.
38. Thummel KE and Wilkinson GR (1998) In vitro and in vivo drug interactions involving human CYP3A. Annu Rev Pharmacol Toxicol 38:389-430.
39. Toft P, Liston AJ, and Viau AY (1973) Synthesis of D4-3-oxo steroids labeled specifically at the 6b-position. J Labelled Compd 9:413-418.
40. Tomaszewski JE, Jerina DM, Levin W, and Conney AH (1976) A highly sensitive radiometric assay for zoxazolamine hydroxylation by liver microsomal cytochrome P-450 and P-448: properties of the membrane-bound and purified reconstituted system. Arch Biochem Biophys 176:788-798.
41. Ueng YF, Kuwabara T, Chun YJ, and Guengerich FP (1997) Cooperativity in oxidations catalyzed by cytochrome P450 3A4. Biochemistry 36:370-381.
42. Wang RW, Newton DJ, Liu N, Atkins WM, and Lu AY (2000) Human cytochrome P-450 3A4: in vitro drug-drug interaction patterns are substrate-dependent. Drug Metab Dispos 28:360-366.
43. Wang RW, Newton DJ, Scheri TD, and Lu AY (1997) Human cytochrome P450 3A4-catalyzed testosterone 6 beta-hydroxylation and erythromycin N-demethylation. Competition during catalysis. Drug Metab Dispos 25:502-507.
44. Waxman DJ, Attisano C, Guengerich FP, and Lapenson DP (1988) Human liver microsomal steroid metabolism: identification of the major microsomal steroid hormone 6 beta-hydroxylase cytochrome P-450 enzyme. Arch Biochem Biophys 263:424-436.
45. Yamazaki H and Shimada T (1997) Progesterone and testosterone hydroxylation by cytochromes P450 2C19, 2C9 and 3A4 in human liver microsomes. Arch Biochem Biophys 346:161-169.
46. Yuan R, Madani S, Wei XX, Reynolds K, and Huang SM (2002) Evaluation of cytochrome P450 probe substrates commonly used by the pharmaceutical industry to study in vitro drug interactions. Drug Metab Dispos 30:1311-1319.
47. Yuan R, Parmelee T, Balian JD, Uppoor RS, Ajayi F, Burnett A, Lesko LJ, and Marroum P (1999) In vitro metabolic interaction studies: experience of the Food and Drug Administration. Clin Pharmacol Ther 66:9-15.
48. Zhang XJ and Thomas PE (1996) Erythromycin as a specific substrate for cytochrome P4503A isozymes and identification of a high-affinity erythromycin N-demethylase in adult female rats. Drug Metab Dispos 24:23-27.