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1. Introduction
2. Goals
3. Key references and disclaimer
4. Reversible modes of enzyme inhibition
5. Enzyme-inhibitor binding equilibria
6. Effects of inhibitors on catalysis steps
7. Inhibition modalities
8. Competitive enzyme inhibitors in clinical use
9. Lovastatin inhibition of HMG-CoA reductase
10. Captopril and enalaprilate inhibition of ACE
11. Competitive inhibitors of HIV protease
12. Noncompetitive enzyme inhibitors in clinical use
13. NNRTI inhibition of HIV reverse transcriptase
14. Uncompetitive enzyme inhibitors in clinical use
15. Methotrexate inhibition of DHFR
16. Dissociation constants: DHFR-ligand complexes
17. DHFR interactions with methotrexate and DHF
18. Inhibition of 5-alpha reductase (1)
19. Inhibition of 5-alpha reductase (2)
20. Dead end inhibitor modality
21. Advantages of reversible inhibition modalities
22. Common non-classical modes of inhibition
23. Enzyme assays for inhibitor discovery
24. Assay criteria for screening
25. Factors that can affect enzyme assays
26. Laboratory measures of enzyme reactions
27. Balanced assay conditions
28. Balanced conditions and physiological context
29. Screening for inhibitors of DHFR (1)
30. Screening for inhibitors of DHFR (2)
31. Measure of signal robustness
32. Hit declaration criteria
33. False positives
34. Promiscuous inhibitors
35. HTS compound follow-up
36. Enzyme inhibitors during lead optimization
37. Compound evaluation flowchart
38. Concentration-response plots
39. Compound IC50 values
40. Effect of substrate on IC50
41. Cheng-Prusoff relationships
42. Hill coefficient
43. Why report Hill coefficients?
44. Biphasic binding
45. Non-ideal 4-parameter fits
46. Non-ideal 2-parameter fits
47. Testing for inhibitor reversibility
48. Defining inhibitor modality and Ki
49. Selectivity in terms of relative Ki values
50. The value of knowing inhibitor modality
51. Suggestions for data reporting
52. Cellular effects and target enzyme inhibition
53. Cellular effects and target engagement
54. Enzyme-cellular correlation plot
55. Summary

Topics Covered

• Enzymes as targets for drug discovery
• Protein-ligand binding equilibria
• Enzyme reaction mechanisms
• Structure and function of enzyme active sites
• Measuring enzyme velocity in the laboratory
• Reversible modes of enzyme inhibition
• Enzyme inhibition as a mechanism of pharmacological intervention in disease
• Clinical value of different inhibition modalities
• Non-classical inhibition
• Enzyme assays and screening criteria
• Lead optimization
• Summary: basic information to look for

Links

Series:

• Small Molecule Drug Discovery

Categories:

• Biochemistry
• Cell Biology
• Diseases, Disorders & Treatments
• Pharmaceutical Sciences

Talk Citation

Publication History

• Published on September 23, 2013

Financial Disclosures

• Prof. Robert Copeland has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.