1. Protein Pharmaceuticals (VI) “Formulation and Delivery”*
07/16/96
Protein Pharmaceuticals (VI) “Formulation and Delivery”
Dr. Aws Alshamsan
Department of Pharmaceutics
Office: AA87
Tel: *

2. Objectives of the Lecture
By the end of this lecture you will be able to:
Describe techniques for optimal protein formulation
Realize the importance of delivery systems for protein pharmaceuticals
Compare different delivery methods

3. Microbial Considerations
Solving the problems
Microbial Considerations
Additives
Storage
Formulation
Delivery

4. Microbial Considerations
Solving the problems
Microbial Considerations
Additives
Storage
Formulation
Delivery

5. Formulation
Biotechnology products are generally sensitve and have considerably short as compared to low-molecular weight drugs.
Optimal formulation is supposed to improve the product stability in vitro and in vivo.

6. Site-Directed Mutagenesis
Bioengineering process that creates a mutation as a defined site in the DNA strand.
Engineered proteins can be tailored for a specific application or improved stability.

7. Site-Directed Mutagenesis

8. Site-Directed Mutagenesis

9. PEGylation
Covalent attachment of PEG to another molecule such as protein.

10. Increase in size to reduce kidney filtration
PEGylation
Significantly increases in vivo half life
Prevents from proteolytic enzymes degradation
Increase in size to reduce kidney filtration
Protein
PEG
Decrease accessibility for proteolytic enzymes and antibodies

11. PEGylated proteins in the market
Brand Name
Drug name
Parent Drug
Indication
Approval Year
Adagen®
Pegadamase
Adenosine deaminase
Severe combined immunodeficiency disease (SCID)
1990
Oncaspar®
Pegaspargase
Asparaginase
Leukaemia
1994
PEG-INTRON®
Peginterferon-a2b
IFN-a2B
Hepatitis C
2000
PEGASYS®
Peginterferon-a2a
IFN-a2A
2001
Neulasta®
Pegfilgrastim
Granulocyte-colony stimulating factor (GCSF)
Neutropenia
2002
Somavert®
Pegvisomant
Growth Hormone antagonist
Acromegaly
2003
Macugen®
Pegaptanib
Anti-VEGFc aptamer
Age-related macular degeneration
2004
Mircera®
Epoetin beta-methoxy polyethylene glycol
Erythropoietin (EPO)
Anemia associate with Kidney disease
2007
Cimzia®
PEG-Certolizumab pegol
Anti-TNF Fab
Rheumatoid arthritis & Crohn’s disease
2008

12. Peptide Micelles Hydrophobic block Hydrophilic block Peptide PEG
CH-CH-CH-CH-CH-CH-CH-CH-CH-CH
| | | | | | | | | |
OH OH OH OH OH OH OH OH OH OH

13. Microbial Considerations
Solving the problems
Microbial Considerations
Additives
Storage
Formulation
Delivery

14. Targeted Micelles

15. Liposomes Phospholipid bilayer Aqueous core 80-100nm in size
Biocompatible
Uni- or Multilamellar
Could be easily functionalized

16. Liposomes Preferential retention in leaky vasculature e.g. cancer
Easily uptaken by macrophages
Overcome by Stealth property (PEGylation)

17. Liposomes

18. Liposomes

19. Microspheres and Nanoparticles
Encapsulation of a protein or peptide inside a porous particle
Constructed of Biodegradable, Biocompatible polymer
PLGA (FDA approved)

20. Microspheres and Nanoparticles

21. Microspheres and Nanoparticles

22. Polymeric Nanoparticle Uptake by Human DCs: SEM image

23. Polymeric Nanoparticle Uptake by Human DCs: Confocal Image

24. Protein Delivery Drugs could be administered by several routs:
Parenteral (injections)
Transdermal
Oral

25. Parenteral

26. Parenteral Delivery Route of delivery for 95% of proteins
Allows rapid and complete absorption
Allows smaller dose size (less waste)
Avoids first pass metabolism
Avoids protein “unfriendly zones”
Problems with overdosing, necrosis
Local tissue reactions/hypersensitivity
Everyone hates getting a needle

27. Oral (GI-MAPS®)

28. Transdermal Patches
Proteins imbedded in a simple matrix with appropriate additives
Patch is coated with small needles that penetrate the dermal layer
Proteins diffuse directly into the blood stream via capillaries
Less painful form of parenteral drug delivery

29. Transdermal patch
Small Pins
Skin

30. Close-up of Patch Pins

31. Exubera (Inhaled Insulin)
Exubera, a dry-powder form of insulin, is inhaled with a special device similar to an asthma inhaler
Exubera normalized blood sugar levels as well as injections did
Patients taking inhaled insulin also reported greater satisfaction and quality of life (for 18+ only)
About 1/5 study subjects developed a mild cough with inhaled insulin
Product pulled in Oct. 2007
Pfizer

32. Oral (Oralin/Oral-Lyn) by GENEREX
It is not oral, it is buccal

33. Oral Insulin (Oralin/Oral-lyn)
Bucchal aerosol delivery system developed by Generex (Approved in Ecaudor and India)

34. Oral Insulin (Oralin/Oral-lyn)
Insulin is absorbed through thin tissue layers in mouth and throat
Insulin is formulated with a variety of additives and stabilizers to prevent denaturation on aerosolization and to stabilize aerosol particles

35. Oral insulin by NOBEX
Conjugate the lysine residue with amphiphilic oligomer
Alkyl chain + PEG
Enhance stability in vivo
Hydrolysis of oligomer release the native structure
Phase II clinical trial (2004)

36. Future Delivery

37. Future Delivery (Biocapsule)
Encapsulation of insulin-secreting cells (insulinoma)
Immune components physically excluded
Glucose molecules pass freely
Insulin is secreted in response to glucose

38. Biocapsules

39. Protein Pharmaceuticals

40. Now you are able to:
Describe techniques for optimal protein formulation
Realize the importance of delivery systems for protein pharmaceuticals
Compare different delivery methods

41. Next Lecture
Biotechnology-based vaccines