VMB 253 Toxicant and Drug Metabolism Schedule of Events-2007:




Jan 4

Introduction, Overview of Drug Metabolism


Jan 9

The P450 Gene Superfamily (nomenclature, favored substrates, inhibitors, inducers)


Jan 11

Chemical Pathways of Drug Metabolism, P450 Catalytic Cycle, Active Site, Substrate binding


Jan 16

Cytochrome P450 Monooxygenase Isozymes, Regio and Stereoselective Metabolism


Jan 18

Inhibitors of the P450 Monooxygenases


Jan 23

Regulation of P450's, and phase II enzymes (XRE, receptors for 2 B isoforms etc); Genetic polymorphisms I


Jan 25

Genetic Polymorphisms II


Jan 30

Hydrolase enzymes (Carboxylesterases, epoxide hydrolases)


Feb 1

Flavin Monooxygenases/N-oxidation


Feb 6

Midterm examination


Feb 8

Conjugation Reactions I. Glucuronyl and Sulfotransferase


Feb 13

Conjugation Reactions II. Glutathione Transferase


Feb 15

Redox cycling/DT Diaphorase/Dihydrodiol dehydrogenase


Feb 20

Methodology in Drug Metabolism-General Approaches, Radiolabeling, Use of Deuterium Isotopes


Feb 22

Considerations in the design and use of metabolism studies in humans and animals-importance of metabolites in toxicities and in toxicity testing


Feb 27

Approaches to Defining Cellular Protein Expression-Immunohistochemistry and quantitative immunoblotting


Van Winkle

March 1

Animal models: Preparation and care of transgenic mice.  Accessing the mutant mouse


March 6

Animal models-continued.  Examples

Lloyd/ Buckpitt

March 8

Recombinant Proteins/Expression systems


March 13

Biomarkers of exposure, metabolism and toxicity


1:30 Friday March 18 but can be changed by class consent.

List of Instructors:
Alan Buckpitt, Molecular Biosciences, 2-7674, Everson Hall 220
Marion Miller, Environmental Toxicology, 2-4526, Meyer Hall
Bruce Hammock
Laura Van Winkle

Course Description:
This course will discuss the general pathways of xenobiotic metabolism in relationship to drug and toxicant interactions with biologic systems. Emphasis will be placed on factors influencing the fate of chemicals and on the techniques used to study various aspects of metabolism. The course attempts to provide up to date information with a window on the future (ie what will happen in the next 5-10 years). Students are encouraged to read selected references which will be provided in the lecture notes. We will attempt to keep one lecture ahead of the presentations in terms of the notes. Please understand that the lectures undergo considerable overhaul each time the course is taught and it may not always be possible to post the notes and power point slides well in advance.

Exams will be essay exams (potentially part take home). Questions will attempt to be integrative and will require understanding of concepts.

Midterm 50%
Final 50%

Prepare a 10 page paper on some aspect of drug metabolism. For example, this might be an excellent way of learning more about the use of recombinant proteins in drug metabolism studies, of the formation of electrophilic metabolites, of genetic differences in metabolism etc. To exercise this option you will need to let me know prior to the first examination. (exams-70%, paper 30%).

Sources of Information:
General Metabolism: **Hayes, A.: Principles and Methods of Toxicology, Fourth Edition, 2000. (The metabolism chapter in this book provides and excellent overview of drug metabolism and the importance of this process in clearance as well as alteration of biologic activity).

Cytochrome P 450: Structure, Mechanism and Biochemistry, Third Edition, Plenum Press, New York, 2005. Discusses structure function relationships, mechanisms of oxygen activation, induction and inhibition of cytochrome P 450 monooxygenases and regulatory mechanisms.

Cytochromes P 450: Metabolic and Toxicological Aspects, CRC press, Boca Raton, 1996. Contains chapters on the role of metabolism and toxicity, on the chemistry of cytochrome P 450 reactions and discusses the P450 families particularly those responsible for metabolism of drugs and toxic substances (families 1-4)

Chapters 7 and 8 from Molecular Toxicology. P. David Josephy and B. Mannervik Oxford University Press, 2006. Pages 204-303 (Worth reading)

Cytochrome P 450 Protocols: Methods in the Molecular Biology, Humana press, New Jersey, 1998. Contains a list of commonly used techniques in the study of cytochrome P. 450 monooxygenase including spectral analysis, purification, reconstitution, catalytic assays, expression of recombinant proteins.

Comprehensive Toxicology, volume 3: Biotransformations. Elsievier press, 1997. Contains overview chapters as well as detailed discussions of each of the major classes of drug metabolizing enzymes. These include the cytochrome P 450 monooxygenases, Flavin containing monooxygenases alcohol and aldehyde dehydrogenase, epoxide hydrolases and phase 2 enzymes. (Unfortunately this volume cannot be purchased separately and is part of a $4000, 13 volume set).

Guengerich FP. Cytochrome P450: what have we learned and what are the future issues? Drug Metab Rev. 2004 May;36(2):159 97. Review

Pharmacogenetics Weinshilboum RM; Otterness DM; Szumlanski CL. Methylation pharmacogenetics: catechol O methyltransferase, thiopurine methyltransferase, and histamine N methyltransferase. Annual Review of Pharmacology and Toxicology, 1999, 39:19 52.

Conjugation Reactions: **Conjugation dependent carcinogenicity and toxicity of foreign compounds. Ed Anders and Dekant. Academic Press, 1994.

Handbook of Drug Metabolism. T Woolf, 1999, very good text, Marcel Dekker

High throughput screening in Drug Metabolism in support of Drug Discovery Annual Rev PTX 40: 133-157, 2000

Combinatorial Chemistry and Contemporary Pharmacology, JPET 293: 705-709, 2000. ** I have these and they may be borrowed for limited use.

Journals in Drug Metabolism

Drug Metabolism and Disposition: (articles pertaining to disposition, metabolic fate, pharmacokinetics, relationship of metabolism to toxicity). Drug Metabolism Reviews: Historical Reviews, proceedings from symposia etc

Xenobiotica: primarily deals with metabolic fate.

Other journals:
Journal of Biological Chemistry, Molecular Pharmacology, etc publish important articles on enzymology, reaction mechanisms, etc.