Effects of PEG conjugation on insulin properties.

The goal of this research was to determine whether the site-specific attachment of poly(ethylene glycol) to insulin could enhance the physical and pharmacological properties of insulin without negatively affecting its biological activity or immunological properties. Electrophilically activated derivatives of low-molecular-weight monomethoxypoly(ethylene glycol) (mPEG) were chemically coupled to insulin via its amino groups at positions phenylalanine-B1 or lysine-B29, with an amide bond being formed between the polymer and protein. The site-specific attachment of mPEG to insulin did not substantially alter insulin's secondary/tertiary structure, self-association behavior, or potency in vivo. However, mPEG attachment did significantly enhance insulin's resistance to aggregation. In addition, the pegylation of insulin almost completely eliminates the resultant conjugate's immunogenicity, allergenicity, and antigenicity. Finally, the conjugates were observed to remain in the systemic circulation for longer periods of time than unmodified insulin after subcutaneous administration.