School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China; Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130000, Jilin, China.
School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China.
Biomaterials. 2014 Jun;35(19):5206-15. doi: 10.1016/j.biomaterials.2014.03.023. Epub 2014 Mar 28.
Fibroblast growth factor 21 (FGF21) is an endocrine-acting hormone that has the potential to treat metabolic diseases, such as type 2 diabetes and obesity. Development of FGF21 into a therapeutic has been hindered due to its low intrinsic bio-stability, propensity towards aggregation and its susceptibility to in vivo proteolytic degradation. In order to address these shortcomings, we've developed recombinant human FGF21 variants by strategically introducing cysteine residues via site-directed mutagenesis, and have also developed a solid-phase nickel affinity PEGylation strategy, whereby engineered, surface-exposed cysteine residues of immobilized proteins were used as a platform to efficiently and site-selectively conjugate with PEG-maleimide. The engineered PEGylated FGF21 conjugates retained its biological functions, as well as demonstrated an increase in half-life by over 211.3 min. By demonstrating the biological activity of the FGF21 analog as a prototype, we have also provided a "generalized" solid-phase approach to effectively increase serum half-life of protein therapeutics.
成纤维细胞生长因子 21（FGF21）是一种具有治疗 2 型糖尿病和肥胖等代谢性疾病潜力的内分泌激素。由于其内在的生物稳定性低、易于聚集以及易在体内发生蛋白水解降解，因此将 FGF21 开发成治疗药物受到了阻碍。为了解决这些缺点，我们通过定点突变策略，在重组人 FGF21 变体中引入半胱氨酸残基，并开发了一种固相镍亲和性聚乙二醇化策略，其中固定化蛋白质的工程化表面暴露的半胱氨酸残基可作为平台，用于有效地和选择性地与聚乙二醇马来酰亚胺缀合。工程化的聚乙二醇化 FGF21 缀合物保留了其生物学功能，并且半衰期延长超过 211.3 分钟。通过展示 FGF21 类似物作为原型的生物活性，我们还提供了一种“通用”的固相方法，可有效延长蛋白治疗药物的血清半衰期。
