College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Republic of Korea; Research Institute of Reference Biolabs. Inc., Seoul, Republic of Korea.
College of Pharmacy and Natural Medicine Research Institute, Mokpo National University, Jeonnam, Republic of Korea.
PLoS One. 2014 May 23;9(5):e96967. doi: 10.1371/journal.pone.0096967. eCollection 2014.
The purpose of this study was to develop a biobetter version of recombinant human interferon-β 1a (rhIFN-β 1a) to improve its biophysical properties, such as aggregation, production and stability, and pharmacokinetic properties without jeopardizing its activity. To achieve this, we introduced additional glycosylation into rhIFN-β 1a via site-directed mutagenesis. Glycoengineering of rhIFN-β 1a resulted in a new molecular entity, termed R27T, which was defined as a rhIFN-β mutein with two N-glycosylation sites at 80th (original site) and at an additional 25th amino acid due to a mutation of Thr for Arg at position 27th of rhIFN-β 1a. Glycoengineering had no effect on rhIFN-β ligand-receptor binding, as no loss of specific activity was observed. R27T showed improved stability and had a reduced propensity for aggregation and an increased half-life. Therefore, hyperglycosylated rhIFN-β could be a biobetter version of rhIFN-β 1a with a potential for use as a drug against multiple sclerosis.
这项研究的目的是开发重组人干扰素-β1a（rhIFN-β1a）的生物改良版本，以改善其聚集、生产和稳定性等生物物理特性，以及药代动力学特性，同时又不影响其活性。为了实现这一目标，我们通过定点突变将额外的糖基化引入 rhIFN-β1a。rhIFN-β1a 的糖基工程导致了一种新的分子实体，称为 R27T，它被定义为 rhIFN-β 无义突变体，在第 80 位（原始位点）和第 27 位 Thr 突变为 Arg 的额外 25 位氨基酸处有两个 N-糖基化位点。糖基工程对 rhIFN-β 配体-受体结合没有影响，因为没有观察到特异性活性的丧失。R27T 表现出更好的稳定性，聚集倾向降低，半衰期延长。因此，高糖基化 rhIFN-β 可能成为 rhIFN-β1a 的生物改良版本，有望用于多发性硬化症的治疗。
