1. PRODUCTION

2. Design of New HIV-Protease Inhibitors and Ritonavir Synthesis

3. BY Sathaporn Prutipanlai Toxicology Program Mahidol University

4. OUT LINE Background Content : Processes of Ritonavir Synthesis : Possibility for synthesis new PIs Conclusion

5. BACKGROUND What is characteristic of HIV Protease enzyme.? HIV Protease is one type of aspartic acid enzyme.

6. Protease enzyme Protease’s function exists as a C 2 - symmetric homodimer. Each monomeric unit contributes one of the conserved catalytic triads(Asp-Thr-gly)

7. HIV Protease Enzyme

8. BACKGROUND (Cont) How does it work.? HIV Protease works by homodimer that cleave gag/pol polypeptide

10. HIV Protease inhibitor development Idea : How are enzyme and substrate interact.? Construct more potent and novel structure of enzyme

11. Protease enzyme

12. Inhibitors represent all three categories Based on peptide isosteres : statin Exploitation the symmetrical properties of the protease dimer. Based on enzyme structure.

13. Type of Designing Protease Inhibitor Non hydrolyzable analog of peptide substrates. Transition-state analogs. Pepstatin-Protease Complex. Two-fold symmetrical or Pseudo symmetrical inhibitor. Structure-Based inhibitor Design

14. Ritonavir Development Peptidomimetic inhibitor Substrate based inhibitor C 2 symmetry-based inhibitor

15. Ritonavir Development Design of C 2 -symmetric inhibitor from an asymmetric substrate compose of 3 steps First: Imposition an axis of symmetry on the peptide functionality in the substrate

16. Ritonavir Development(cont) Second: Arbitary deletion of either the N-terminal or C-terminal. Third: C 2 symmetry operation is applied to the remaining portion to generate a symmetric core unit.

17. C 2 -symmetric inhibitor to HIV protease enzyme Imposition of C 2 -symmetry axes on an asymmetric substrate or inhibitor Kempf, D.J. et.al. 1993

19. C 2 Symmetric HIV PIs Kempf, D.J. et.al 1993

20. Synthesis of Symmetric Inhibitor Core Units Three general categories. : Linear, nonsymmetric syntheses : Symmetric combination of identical halves : Bifunctionalization of a C 2 symmetric precursor

21. Linear, Nonsymmetric syntheses Kempf,D.J. 1994

22. Synthesis of Symmetric Inhibitor Core Units(cont) Three general categories. : Symmetric combination of identical halves : Bifunctionalization of a C 2 symmetric precursor

23. Effect of C 2 -symmetric inhibitors to HIV protease

25. Processes of Ritonavir Synthesis

26. +  -aminoaldehyde Scheme 1 vcl 3 Zn 2-5-bis-N- ((benzyl)oxy)carbonyl)amino-3, 4-diacydroxy-1, 6 diphenylhexane (diols) Patent#5,846,987

27. Scheme 2 2-5-bis-N- ((benzyl)oxy)carbonyl) amino-3, 4-diacydroxy- 1, 6 diphenylhexane bromoacetate Hydrolysis Cyclization Reduction 2-5-bis-N-((benzyl)oxy)carbonyl)amino)1, 6 diphenyl-3-hydroxyhexane Patent#5,846,987 2,5-Bis-(N(((benzyl)oxyl)carbonyl amino)-3-4-epoxy-1,6 diphenylhaxane

28. Scheme 2 (cont) 2-5-bis-N-((benzyl)oxy)carbonyl)amino)1, 6 diphenyl-3-hydroxyhexane hydrolysis 2, 5, -diamino-1, 6diphenyl- 3-hydroxyhexane Patent#5,846,987

29. Scheme 3 acylation Patent#5,846,987 2,5,-Diamino-1,6- diphenyl-3- hydroxyhexane 6(1-Amino-2- phenyl)-4-benzyl- 2-phenyl-3-aza-2- boro-1- oxacyclohexane 5-(Thiazolyl)methyl)-(4- nitrophenyl)carbamate 5-Amino-2-(N-((5-thiazolyl)methoxy carbonyl)amino)-1,6-diphenyl-3- hydroxyhexane

30. Scheme 3 (cont) Patent#5,846,987 5-Amino-2-(N-((5- thiazolyl)methoxy carbonyl)amino)-1,6-diphenyl-3- hydroxyhexane Coupling reaction Ritonavir N-((N-methyl-N-((2-isopropyl-4-thiazoyl methyl)amino)carbonyl-L-valine

31. Molecular structure of Ritonavir

33. Ritonavir and protease enzyme

34. Indinavir and Protease Enzyme

35. Indinavir Saquinavir Ritonavir Nelfinavir

36. Possibility to design new protease inhibitor Factor : Hydrophobic : High oral bioavilability : Low hepatic clearance : Low toxicity

37. CONCLUSION Factor that influence drug design : Pharmacokinetic : Pharmacodynamic : Interaction between inhibitors and HIV- protease enzyme

38. THANK YOU Dr. Poonsak Dr. Maria Kartalou Dr. Suwit CRI’s Staff YOUR ATTENTION

39. THE END