Cytochrome P450: In Vitro Methods and Protocols (Methods in Pharmacology and

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1. Cytochrome P450: In Vitro Methods and Protocols

Zhengyin Yan and Gary W. Caldwell

2. Characteristics of Major Drug Metabolizing Cytochrome P450 Enzymes

Kevin Johnson, Dian Su, and Donglu Zhang

3. Quantitative Determination of Cytochrome P450 Using LC/MS/MS

Ryan H. Takahashi, Brahim Achour, and Bhagwat Prasad

4. Spectral Quantitation of Total Cytochrome P450 in Microsomes Using a Single-Beam Spectrophotometer

David M. Stresser

5. Cytochrome P450 Enzyme Kinetics: Km and Vmax Determinations Using Metabolite Formation and Substrate Depletion Methods

Jing Wang

6. Cytochrome P450 Inhibition Assay Using Human Liver Microsomes

Jia Jia

7. Evaluation of Time-Dependent Cytochrome P450 Inhibition Using the Area Under the Curve Shift Method

Julie Huang

8. Assessing Cytochrome P450 Time Dependent Inhibition (IC50 Shift Assay) Using Both Diluted and Non-Diluted Methods

Yu Wang, Lili Xie, and Jing Ni

9. Cytochrome P450 Gene Regulation: Reporter Assays to Assess Pregnane X Receptor (PXR, NR1I2) Activation

Marija Pinne and Judy L. Raucy

10. Cytochrome P450 Gene Regulation: Reporter Assays to Assess Aryl Hydrocarbon Receptor (HLHE76, AhR) Activation and Antagonism

Marija Pinne and Judy L. Raucy

11. Cytochrome P450 Gene Regulation: Reporter Assays to Assess Direct and Indirect Activation of Constitutive Androstane Receptor (CAR, NR1I3)

Marija Pinne and Judy L. Raucy

12. Determination of In Vitro Cytochrome P450 Induction Potential Using Cryopreserved Human Hepatocytes

Susan Wong

13. Assessing Cytochrome P450 3A (CYP3A) Induction and Suppression Using Primary Human Hepatocyte Spheroids

Jing Wang and Eugene Chen

14. Cytochrome P450-Mediated Metabolic Stability Assay in Liver Microsomes

Pasquale Carione

15. Assessment of Cytochrome P450 Metabolic Clearance Using Hepatocyte Suspension

Jingwei Cai and Hadil Shalan

16. Cytochrome P450 Knock-Out Assay with Non-Selective Inhibitors

Pasquale Carione and Zhengyin Yan

17. Differentiation of Cytochrome P450-Mediated from Non-CYP-Mediated Metabolism: Aldehyde Oxidase and Xanthine Oxidoreductase

Kirk D. Kozminski and Michael A. Zientek

18. In Vitro Reaction Phenotyping of Cytochrome P450 Enzymes

Susan Wong and Zhengyin Yan

19. Cytochrome P450-Mediated Drug Bioactivation Assay: An Untargeted High Resolution Accurate Mass LC/MS Assay

Kevin M. Johnson

20. Simultaneously Assessing Concentration Changes in 20 Biochemical Pathways as a Result of Drug Dosing and Cytochrome P450 and Non-Cytochrome P450-Mediated Metabolism: An Untargeted Metabolomics GC/MS Assay

Yanping Lin, Ying Li, and Gary W. Caldwell

21. Simultaneously Assessing Concentration Changes in 17 Biochemical Pathways as a Result of Drug Dosing and Cytochrome P450 and Non-Cytochrome P450-Mediated Metabolism: A Quasi-Untargeted Metabolomics LC/MS Assay

John A. Masucci, Feng Liang, Kerem Bingol, Vince Windisch, and Gary W. Caldwell

22. Assessing Amino Acid Concentration Changes as a Result of Drug Dosing and Cytochrome P450 and Non-Cytochrome P450-Mediated Metabolism

Kerem Bingol, John A. Masucci, and Gary W. Caldwell