Yin Jun, Bao Lichen, Tian Hong, Wang Qun, Gao Xiangdong, Yao Wenbing
State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, Nanjing, China.
Br J Pharmacol. 2016 Jul;173(14):2208-23. doi: 10.1111/bph.13499. Epub 2016 Jun 3.
Chemical conjugation of therapeutic proteins with polyethylene glycol (PEG) is an established strategy to extend their biological half-life (t1/2 ) to a clinically useful range. We developed a novel uncharged and unstructured recombinant polypeptide composed of five amino acids (P, S, T, A and G), named PsTag, as another approach to extend the t1/2 of human FGF21, with increased hydrodynamic radius.
Human FGF21 was fused with PsTag polymers of differing lengths (200 - 600 residues). Three fusion proteins and native FGF21 were produced in Escherichia coli. The biophysical characteristics, metabolic stability, immunogenicity and pharmacokinetics in were assessed in first. In lean and diet-induced obese (DIO) mice, effects on body weight, oral glucose tolerance tests and levels of relevant hormones and metabolites were studied.
Fusion proteins were solubly expressed in E. coli and prolonged the t1/2 from 0.34h up to 12.9 h in mice. Fusion proteins were also biodegradable, thus avoiding vacuole formation, while lacking immunogenicity in mice. In DIO mice, administration of PsTag fused to FGF21 reduced body weight, blood glucose and lipids levels and reversed hepatic steatosis.
The novel recombinant polypeptide, PsTag, should be useful in the development of biological drugs with properties comparable to those achievable by PEGylation, but with potentially less side effects. In mice, fusion of FGF21 to PsTag prolonged and potentiated pharmacological effects of native FGF21, and may offer greater therapeutic effects in treatment of obesity.
将治疗性蛋白质与聚乙二醇（PEG）进行化学偶联是一种已确立的策略，可将其生物半衰期（t1/2）延长至临床有用范围。我们开发了一种由五个氨基酸（P、S、T、A和G）组成的新型不带电荷且无结构的重组多肽，命名为PsTag，作为延长人FGF21的t1/2的另一种方法，同时增加流体力学半径。
将人FGF21与不同长度（200 - 600个残基）的PsTag聚合物融合。在大肠杆菌中生产了三种融合蛋白和天然FGF21。首先评估了它们的生物物理特性、代谢稳定性、免疫原性和药代动力学。在瘦小鼠和饮食诱导的肥胖（DIO）小鼠中，研究了对体重、口服葡萄糖耐量试验以及相关激素和代谢物水平的影响。
融合蛋白在大肠杆菌中可溶表达，并将小鼠体内的t1/2从0.34小时延长至12.9小时。融合蛋白也具有生物可降解性，从而避免了液泡形成，同时在小鼠中缺乏免疫原性。在DIO小鼠中，给予与FGF21融合的PsTag可降低体重、血糖和血脂水平，并逆转肝脂肪变性。
新型重组多肽PsTag在开发具有与聚乙二醇化相当性质但潜在副作用可能更少的生物药物方面应具有实用性。在小鼠中，将FGF21与PsTag融合可延长并增强天然FGF21的药理作用，并且在肥胖治疗中可能提供更大的治疗效果。
