1. Protein Therapeutics: Delivery Devin Hudson

2. Delivery Methods Intravenously Subcutaneously Suppository Intranasally Orally * Transinfection *

3. Liquid Filled Nanoparticles as a Drug Delivery Tool. Venkatesan, N., Yoshimitsu, J., Ito, Y., Shibata, N. Takada, K. (2005) Biomaterials. 26, 7154-7163.

4. Barriers to Oral Bioavailability Percent Bioavailability: BA% Poor Membrane Permeability Enzymatic Degradation Past Attempts: liposomes, nanoparticles, micro-spheres, hydragels, mucoadhesives, micro-emulsions

5. Concept

6. Erythropoietin (EPO) 1BUY PDB: 1BUY Hormone: Produced in Kidneys Erythropoiesis

7. Porous Absorbents Carbon Nano Tube Carbon Nanohorn Fulleren es Other Porous Substrates: Silicon Dioxide, Charcoal, bamboo charcoal

8. Treatment Jejunum: large surface area Blood EPO measured via Jugular vein with ELISA

9. Formulation G: labrasol Absorption Enhancers

10. Formulation A: CNT Porous Absorbents

11. Formulation G: Casein Protease Inhibitors casein vs lactoferrin

12. Conclusion BA% = 11.5

13. Systemic Delivery of Secreated Protein by Grafts of Epidermal Keratinocyctes: Prospects for Keratinocyte Gene Therapy. Fenjves, E. S., Smith, J., Zaradic, S. and Lorne, B. T. (1994) Human Gene Therapy. 5, 1241-1248 Wikipedia.org

14. Apolipoprotein E (apoE) PDB: 1B68 Apolipoprotein Essential for the normal catabolism of triglyceride-rich lipoprotein constituents Chylomicrons Transport dietary lipids from the intestines to other locations in the body C C 34kD Deficiency related to familiar heart disease Wikipedia

15. Concept Transfect with recombinant apoE-HA1 (pSV2neo) Graft skin ELISA: Two Tests Wikipedia.org

16. Keratinocyctes and Aythmic Mice Stable Long Term Grafts Effectively Transduced via retrovirus Secretory tissue Newborn Foreskin Nude Mouse Immune- deficient No Rejection Response Xenografts Wikipedia

17. Fractionation – Ficoll 400 Non-fractionated: total Small: basal compartment rich Intermediate: poorly characterized Large: terminally differentiated suprabasal cells Basal compartment keratinocyctes excrete endogenous apoE Mature Differentiated keratinocyctes do not excrete endogenous apoE Recomibant apoE expressed by any transfected keratinocytes

18. Long Term Success Blood Serum apoE after 28 days Previous work: Viral promoters shut off over time

19. Conclusion Higher Recombinant Expression in vivo Expansion in suprabasal cells in grafts Estimated that graft covering 2% of human skin could provide 6.5-8.9mg of recombinant protein per day.

20. Oral vs Graft Pros: Easily Manage Dosing Quit/Start/Change Treatment Minimally Invasive Lower Short Term Expense Cons: Require On-going Care Suffer from Varied Absorption Pros: Doesn’t Require On-Going Care Lower Long Term Expense Less Margin for Patient Error Less Risky than Systemic Transfection Cons: Invasive Treatment Can Not be Stopped Easily

21. Refrences Skin Patch Fenjves, E. S., Smith, J., Zaradic, S. and Lorne, B. T. (1994) Systemic Delivery of Secreated Protein by Grafts of Epidermal Keratinocyctes: Prospects for Keratinocyte Gene Therapy. Human Gene Therapy. 5, 1241-1248. Review Paper Leader, B., Baca, Q, J,. and Golan, D, E. (2008) Protein therapeutics: a summary and pharmacological classification. Nature Publishing Group. 7, 21-39. Oral Venkatesan, N., Yoshimitsu, J., Ito, Y., Shibata, N. Takada, K. (2005) Liquid filled nanoparticles as a drug delivery tool. Biomaterials. 26, 7154-7163.