Site-specific attachment of functionalized poly(ethylene glycol) to the amino terminus of proteins.
A convenient method for the construction of site-specifically modified poly(ethylene glycol)-protein conjugates is described. This method relies on the ability to generate a reactive carbonyl group in place of the terminal amino group. If the protein has N-terminal serine or threonine, this can be done by very mild periodate oxidation and generates a glyoxylyl group. A method less restricted by the nature of the N-terminal residue, but which requires somewhat harsher conditions, is metal-catalyzed transamination, which gives a keto group. The N-terminal-introduced reactive carbonyl group specifically reacts, under mild acidic conditions, with an aminooxy-functionalized poly(ethylene glycol) to form a stable oxime bond. Using polymers of different size and shape (linear or multibranched), various conjugates of IL-8, G-CSF, and IL-1ra were constructed and further characterized with respect to their biological activity and pharmacokinetic behavior in rats. Unlike most previous methods, this approach places a single PEG chain at a defined site on the protein. It should therefore be more likely to conserve biological activity when the latter depends on interaction with another macromolecule (unlike enzymic activity which often survives multiple PEGylation).
Gaertner
H F
HF
Département de Biochimie Médicale, Centre Médical Universitaire, Geneva, Switzerland.
Offord
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RE
D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D015232:Dinoprostone / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000493:pharmacokinetics / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D006801:Humans; D007202:Indicators and Reagents; D053590:Interleukin 1 Receptor Antagonist Protein; D016209:Interleukin-8 / Q000493:pharmacokinetics / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology; D010446:Peptide Fragments / Q000737:chemistry*; D011092:Polyethylene Glycols; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D051381:Rats; D012694:Serine; D012795:Sialoglycoproteins / Q000493:pharmacokinetics / Q000494:pharmacology; D012867:Skin / Q000187:drug effects / Q000378:metabolism; D013329:Structure-Activity Relationship; D013912:Threonine
C507763:IL1RN protein, human; D007202:Indicators and Reagents; D053590:Interleukin 1 Receptor Antagonist Protein; D016209:Interleukin-8; D010446:Peptide Fragments; D011506:Proteins; D012795:Sialoglycoproteins; D016179:Granulocyte Colony-Stimulating Factor; D013912:Threonine; D011092:Polyethylene Glycols; D012694:Serine; D015232:Dinoprostone