Jo Yeong Woo, Youn Yu Seok, Lee Sung Hee, Kim Byong Moon, Kang Soo Hyung, Yoo Moohi, Choi Eung Chil, Lee Kang Choon
Research Laboratories, Dong-A Pharm. Co., Ltd., Yongin 449-905, Republic of Korea.
Int J Pharm. 2006 Feb 17;309(1-2):87-93. doi: 10.1016/j.ijpharm.2005.11.035. Epub 2006 Jan 9.
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.
聚乙二醇（PEG）尺寸和形状的恰当选择是聚乙二醇化技术中最重要的要点之一。因此，已广泛开发出各种尺寸和形状的PEG以赋予特定性能。在本研究中，一种独特的三聚体结构、43 kDa的PEG通过形成酰胺键与α-2a干扰素（IFN）偶联，以改善药代动力学性质并使IFN生物活性的损失最小化。利用这种独特的PEG制备的单聚乙二醇化IFN（PEG(3)-IFN）通过阳离子交换色谱法和基质辅助激光解吸电离飞行时间质谱法进行纯化和表征。对PEG(3)-IFN的体外生物活性、体内稳定性和药代动力学进行了研究，并与天然IFN进行了比较。PEG(3)-IFN表现出与天然IFN相当的体外生物活性，并且在皮下注射后在大鼠血清和各种器官中偶联连接具有优异的稳定性。此外，它在大鼠体内吸收缓慢且清除率显著降低，从而使生物半衰期相比天然IFN增加了约40倍。这是关于将独特的三聚体结构PEG应用于生物活性蛋白的首次报道。结果表明，独特的三聚体结构43 kDa PEG可为改善药代动力学性质以及在临床应用中维持治疗性蛋白的生物活性提供一些优势。
