PEGylation: A successful Approach to Drug Delivery

Presentation By: Ryan Mulkeen

Why is PEGylation a Hot Topic

First described in the 1970s, An important development as PEGylation modifies a protein extensively with maintenance of activity Non-toxic, non-immunogenic, highly soluble in water and FDA approved Since 1990 many PEGylated drugs have been synthesized and approved including drugs for cancer, Hepatitis, HIV, and MS Low cost of manufacturing Part of a multi-billion dollar molecular medicines market
Chemical & Engineering News 83 (2005) 21-29 PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G.

Current Drug Problems

Diagram: transpeg.pbwiki.com

PEG - Polyethylene Glycol

Synthesized from the polymerization of ethylene oxide Using chemical tools to link PEG molecules to native proteins can yield conjugates with more favorable behavior
Diagram: transpeg.pbwiki.com

Major Advantages to PEGylated Molecules

PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G.

Reducing Kidney Filtration

PEGylation significantly increases the apparent size of the conjugated drug compound

Diagram: transpeg.pbwiki.com

PEG is not ready for conjugation reactions by itself

1.Needs a capped terminus with unreactive moiety 2. Other end has reactive moiety that is covalently with reactive partner (protein, peptide, other compounds) Diagram: transpeg.pbwiki.com

Paradigm of in vivo PEG linkage

Cleavage to proceed without liberating any reactive and potentially toxic side products

Diagram: transpeg.pbwiki.com

Derivatives

PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G.

Derivatives

PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G.

Derivatives

PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G.

Derivatives

PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G.

Limitations in PEGylation
Isomerization of polymer Excretion from the body Polydispersity

Limitations: Polydispersity

Data of a 5 kDa polymer synthesis

Peptide and protein PEGylation: a review of problems and solutions. Biomaterials. 2001; 22 ,Veronese M.

Interferon--1b
Drug used for treatment of MS (multiple sclerosis Short half-lives Human antibodies can further decrease bioavailability

Image: www.multiplesklerosemailingliste.info

Goals
After expressing in E.Coli: Lower Aggregation/ Prolong Solubility Lower Immunogenicity Improve Potency Approach: Conjugate with varying PEG sizes and # of conjugated sites (mono and multi PEGylation)

Aggregation Studies

Aggregation forms as pH rises Measured by HPLC

StructureFunction Engineering of Interferon-Beta-1b for Improving Stability, Solubility, Potency, Immunogenicity, and Pharmacokinetic Properties by SiteSelective Mono-Pegylation. Bioconjugate Chem. 2006; 17, 618-630. Basu A. et al.

Immunogenicity

Assay detecting antigens in a sample IgG high affinity for drug without PEG
StructureFunction Engineering of Interferon-Beta-1b for Improving Stability, Solubility, Potency, Immunogenicity, and Pharmacokinetic Properties by Site-Selective Mono-Pegylation. Bioconjugate Chem. 2006; 17, 618-630. Basu A. et al.

Measurement once per week dosing per week

Pharmacokinetic Studies

StructureFunction Engineering of Interferon-Beta-1b for Improving Stability, Solubility, Potency, Immunogenicity, and Pharmacokinetic Properties by Site-Selective Mono-Pegylation. Bioconjugate Chem. 2006; 17, 618-630. Basu A. et al.

Pharmacokinetic Studies

StructureFunction Engineering of Interferon-Beta-1b for Improving Stability, Solubility, Potency, Immunogenicity, and Pharmacokinetic Properties by Site-Selective Mono-Pegylation. Bioconjugate Chem. 2006; 17, 618-630. Basu A. et al.

Future questions for PEGylation

What can we learn from the binding/targeting chemistry? Drugs currently on the market. Can they be improved?

References
PEGylation, successful approach to drug delivery. Drug Discovery Today. 2005 Nov; 10(21) Veronese M. and Pasut G. Peptide and protein PEGylation: a review of problems and solutions. Biomaterials. 2001; 22 ,Veronese M. StructureFunction Engineering of Interferon-Beta-1b for Improving Stability, Solubility, Potency, Immunogenicity, and Pharmacokinetic Properties by Site-Selective Mono-Pegylation. Bioconjugate Chem. 2006; 17, 618-630. Basu A. et al. Chemical & Engineering News 83 (2005) 21-29 GlycoPEGylation of recombinant theraputic proteins produced in Escherichia coli. Glycobiology. 2006; 16(9) Defrees et.al.

Extra Slide: A small approved drug list