Glycans in the Biotechnology and Pharmaceutical Industries Lecture 42 Carolyn R. Bertozzi

1. Glycans in the Biotechnology and Pharmaceutical Industries Lecture 42 Carolyn R. Bertozzi UC Berkeley

2. Lecture Outline • Examples of approved carbohydrate-based drugs • Development of drugs to treat influenza • Development of selectin inhibitors for inflammation • Imino-sugars as drug candidates for storage diseases

3. Examples of approved carbohydrate drugs: Substance Indication Company Acarbose Diabetes Bayer AG AMVISC Opthalamic surgery Anika Therapeutics Hyalgan Osteoarthritis FIDIA/Sanofi Lovenox Cardiovascular disease Aventis Miglitol Diabetes Bayer AG ORTHOVISC Osteoarthritis Anika Therapeutics Relenza Influenza Glaxo Smithkline Tamiflu Influenza Roche SOLARASE Actinic keratosis Hyal Pharmaceuticals Topamax Epilepsy J & J Voglibose Diabetes Takedo/Abbott

4. Examples of carbohydrate-based drugs

5. Examples of glycosylated natural products

6. From: Hudson, P. J.; Souriau, C. Nature Medicine2003, 9, 129-134 Examples of approved glycoprotein drugs: Substance Company Erythropoietin Amgen, J&J Tissue plasminogen activator Genentech Interleukin-2 Chiron Cerezyme Genzyme Monoclonal antibodies Many

7. Lecture Outline • Examples of approved carbohydrate-based drugs • Development of drugs to treat influenza • Development of selectin inhibitors for inflammation • Imino-sugars as drug candidates for storage diseases

8. bacterium virus cell toxin hormone Cell surface oligosaccharides are determinants of cell recognition glycoprotein

9. Microbial pathogens bind to cell surface sugars as a first step during infection Influenza virus - “Flu” HIV - AIDS Helicobacter pylori - Ulcers Escherichia coli - Meningitis Pseudomonas aeruginosa - Pneumonia Trypanosomes - African sleeping sickness Plasmodium falciparum - Malaria

10. The influenza virus has two membrane-associated proteins, hemagglutinin and neuraminidase Neuraminidase cleaves sialic acid (enzyme) Hemagglutinin binds sialic acid (receptor) Sialic acid (SA) Host cell

11. Neuraminidase cleaves sialic acid and liberates new virus New viruses assemble at membrane Endocytosis Membrane fusion and release of viral particle Replication Life cycle of the influenza virus SA Hemagglutinin binds sialic acid to initiate infection SA SA SA Host cell SA SA

12. influenza virus Sialic acid analogs block influenza virus infection Sialic acid Cell “C-Glycoside” of sialic acid

13. Ea (uncat) Ea (cat) Transition state analogs are potent enzyme inhibitors Enzymes catalyze reactions by preferential binding of the transition state vs the ground state

15. Planarity (sp2) 2,3-Anhydro sialic acid Ki = 10-6 M Also inhibits human neuraminidase Design of transition state analog neuraminidase inhibitors ‡

16. Binding pocket of Flu neuraminidase based on X-ray structure Relenza (Glaxo Smithkline) Ki = 10-10 M Tamiflu (Hoffman La Roche) Ki = 10-10 M Structure-based design of more potent and selective neuraminidase inhibitors

17. Lecture Outline • Examples of approved carbohydrate-based drugs • Development of drugs to treat influenza • Development of selectin inhibitors for inflammation • Imino-sugars as drug candidates for storage diseases

18. A hallmark of inflammation is the recruitment of leukocytes from the bloodstream into surrounding tissues Tissue Blood vessel Leukocyte Activated endothelium

19. The initial attachment of leukocytes to endothelial cells at sites of inflammation is mediated by the selectins Leukocyte L-selectin E-selectin P-selectin Endothelial cell

20. Inflammatory diseases involving the selectins: 1. Rheumatoid arthritis 2. Asthma 3. Transplant rejection 4. Psoriasis 5. Inflammatory bowel disease 6. Ischemia/reperfusion injury 7. Diabetes 8. Multiple sclerosis 9. Many more….. Inhibitors of selectin-mediated cell adhesion would be broad spectrum anti-inflammatory agents

23. Limitations of sLex as a therapeutic agent • Lack of potency • Poor pharmacokinetics • Difficult and expensive synthesis Possible solutions • Glycomimetics • Oligomerization • Glycoprotein constructs

26. (in clinical development at Texas Biotech.)

27. Limitations of sLex as a therapeutic agent • Lack of potency • Poor pharmacokinetics • Difficult and expensive synthesis Possible solutions • Glycomimetics • Oligomerization • Glycoprotein constructs

29. Nagy and coworkers

30. Kiessling and coworkers

31. Limitations of sLex as a therapeutic agent • Lack of potency • Poor pharmacokinetics • Difficult and expensive synthesis Possible solutions • Glycomimetics • Oligomerization • Glycoprotein constructs

32. Approaches to selectin inhibitors inspired by the discovery of their biological ligands

33. Structure of the biological selectin ligands

34. Determinants of PSGL-1 required for P-selectin binding

35. A potent P-selectin inhibitor based on PSGL-1:Recombinant PSGL-Ig (TS-1) 47 N-terminal residues from PSGL-1 Human IgG1 Fc (inactivated) Features: • Good potency (nM Kd) • Good PK (serum 1/2-life = 2-3 wks) • Also inhibits L-selectin

36. Leukocyte adhesion to endothelial cells is mediated by L-selectin binding to sulfated sialyl Lewis x Leukocyte 6-Sulfo sialyl Lewis x L-Selectin { Endothelial cell

38. Lecture Outline • Examples of approved carbohydrate-based drugs • Development of drugs to treat influenza • Development of selectin inhibitors for inflammation • Imino-sugars as drug candidates for storage diseases

39. Defects in glycolipid degradation lead to lysosomal storage disorders From: Dwek, R. A., et al.Nature Rev. Drug Disc. 2001, 1, 65-75.

40. Imino-sugars can block biosynthesis of glycolipid precursors

42. Numerous companies have been foundedon glycobiology platforms