Atypical kinetic profiles observed with drug metabolizing enzymes, such as the cytochromes P450, can confound in vitro determinations of kinetic parameter estimates. During the development of new drug entities, inaccurate kinetic parameter estimates can lead to incorrect decisions as to the potential success of the drug compound. The long term goals of this research are to understand the biochemical mechanisms resulting in atypical kinetic profiles and develop computer models capable of predicting the occurrence of atypical kinetics. It is generally accepted that two (or more) substrate molecules can bind within the active site of cytochrome P450 enzymes and this phenomenon plays a role in atypical kinetics. NMR work from our laboratory has demonstrated two molecules present in the active site and the presence of an effector causes movement of the primary substrate closer to the heme. Increases in P450 reaction efficiency and reductions in reaction cycle uncoupling also have been observed during CYP2C9 heteroactivation, but preliminary data suggest that not all activator compounds act by the same mechanism. The structure-activity basis for these differential effects is unknown. More recently, we have observed that genetic variants of CYP2C9 exhibit greater degrees of heteroactivation than wild-type enzyme, suggesting that enzyme conformation may also be a factor in the observation of atypical kinetics. The current competing renewal builds upon the above findings and seeks to ascertain; a) the mechanism(s) by which effector molecules alter P450 reaction cycle efficiency and the structure-activity relationships of molecules causing these effects, b) the changes in substrate-heme iron distances and substrate orientation produced by different effectors, and c) how enzyme amino acid alterations affect substrate/effector-protein interactions and thus, activation. From these data, computer models will be developed to predict the occurrence of atypical kinetics and potential drug interactions to assist in the drug development process.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM063215-05
Application #
7061796
Study Section
Xenobiotic and Nutrient Disposition and Action Study Section (XNDA)
Program Officer
Okita, Richard T
Project Start
2001-07-01
Project End
2009-03-31
Budget Start
2006-05-01
Budget End
2007-04-30
Support Year
5
Fiscal Year
2006
Total Cost
$238,098
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Zhou, Jin; Tracy, Timothy S; Remmel, Rory P (2011) Correlation between bilirubin glucuronidation and estradiol-3-gluronidation in the presence of model UDP-glucuronosyltransferase 1A1 substrates/inhibitors. Drug Metab Dispos 39:322-9
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Zhou, Jin; Tracy, Timothy S; Remmel, Rory P (2010) Bilirubin glucuronidation revisited: proper assay conditions to estimate enzyme kinetics with recombinant UGT1A1. Drug Metab Dispos 38:1907-11
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