Long-acting interferon-alpha 2a modified with a trimer-structured polyethylene glycol: preparation, in vitro bioactivity, in vivo stability and pharmacokinetics

• Published in: International journal of pharmaceutics 2006-02, Vol.309 (1-2), p.87-93
• Main Authors: Jo, Yeong Woo, Youn, Yu Seok, Lee, Sung Hee, Kim, Byong Moon, Kang, Soo Hyung, Yoo, Moohi, Choi, Eung Chil, Lee, Kang Choon
• Format: Article
• Language: English
• Subjects: Animals; Antiviral Agents - chemical synthesis; Antiviral Agents - pharmacokinetics; Antiviral Agents - pharmacology; Cell Line; Cell Proliferation - drug effects; Chemistry, Pharmaceutical; Delayed-Action Preparations; Dogs; Drug Stability; Interferon-alpha - chemical synthesis; Interferon-alpha - pharmacokinetics; Interferon-alpha - pharmacology; Microbial Sensitivity Tests; Molecular Weight; Polyethylene Glycols - chemical synthesis; Polyethylene Glycols - pharmacokinetics; Polyethylene Glycols - pharmacology; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Technology, Pharmaceutical - methods; Vesicular stomatitis Indiana virus - drug effects; Vesicular stomatitis Indiana virus - growth & development
• ISSN: 0378-5173
• DOI: 10.1016/j.ijpharm.2005.11.035

The proper selection of size and shape for polyethylene glycol (PEG) is one of the most important points in PEGylation technology. Therefore, PEGs of various sizes and shapes have been widely developed to endow specific properties. In this study, a unique, trimer-structured, 43 kDa PEG was conjugated to interferon-alpha 2a (IFN) by forming an amide bond to improve the pharmacokinetic properties and minimize the loss of IFN bioactivity. Mono-PEGylated IFN (PEG(3)-IFN) prepared by utilizing this unique PEG was purified and characterized by cation-exchange chromatography and MALDI-TOF mass spectrometry. The in vitro bioactivity, in vivo stability, and pharmacokinetics of PEG(3)-IFN were examined and compared to those of native IFN. PEG(3)-IFN exhibited comparable in vitro bioactivities to native IFN and an excellent stability of the conjugation linkage in rat serum and various organs following subcutaneous injection. Furthermore, it showed slow absorption and markedly reduced clearance in rats, thereby increasing the biological half-life by about 40-fold compared to that of native IFN. This is the first report on the application of unique, trimer-structured PEG to bioactive proteins. The results suggest that unique, trimer-structured 43 kDa PEG can provide some advantages to improve the pharmacokinetic properties and to maintain the bioactivity of therapeutic proteins in clinical use.