Prolonging the Action of Protein and Peptide Drugs by a Novel Approach of Reversible Polyethylene Glycol Modification

• Published in: The Journal of biological chemistry 2004-09, Vol.279 (37), p.38118-38124
• Main Authors: Tsubery, Haim, Mironchik, Marina, Fridkin, Mati, Shechter, Yoram
• Format: Article
• Language: English
• Subjects: Anhydrides - pharmacology; Animals; Blood Glucose - metabolism; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Exenatide; Fluorenes - chemical synthesis; Fluorenes - pharmacology; Glucose - metabolism; Human Growth Hormone - chemistry; Humans; Hydrogen-Ion Concentration; Hydrolysis; Maleimides - chemical synthesis; Maleimides - pharmacology; Mice; Models, Chemical; Peptides - chemistry; Peptides - pharmacology; Polyethylene Glycols - chemistry; Propionates - pharmacology; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectrophotometry; Succinimides - pharmacology; Temperature; Time Factors; Ultraviolet Rays; Venoms - chemistry
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M405155200

Polyethylene glycol (PEG)-conjugated therapeutic peptides/proteins have been shown to exhibit clinical properties superior to those of their corresponding unmodified parent molecules. However, the desirable pharmacological features gained by protein PEGylation become irrelevant if conjugates are inactivated or cannot reach their target tissues. Here we describe the design and synthesis of MAL-FMS-OSU. This bifunctional agent enables PEG chains to be linked to peptides and proteins through a slowly hydrolysable chemical bond. PEG-FMS-peptide/protein conjugates thus formed undergo spontaneous hydrolysis at a slow rate upon incubation at pH 8.5, 37 °C with a t½ value of 8–14 ± 2 h, generating the unmodified parent molecule. The validity of this approach was studied with exendin-4 and human growth hormone. A single subcutaneous administration of PEG40,000-FMS-exendin-4 facilitated a prolonged and stable reduction in glucose levels in mice (t½ = 30 ± 2 h) and exceeded the effect obtained by the same dose of the native hormone by 7–8 times.