Department of Chemical Engineering, Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India.
AAPS J. 2017 Nov;19(6):1826-1841. doi: 10.1208/s12248-017-0139-8. Epub 2017 Sep 5.
Estimating impact of the various product-related variants and impurities on a biotherapeutic's safety and efficacy is an essential requirement in the quality by design paradigm. In view of the limited role that clinical studies offer in this regard, we demonstrate a preclinical approach to achieve this for granulocyte colony-stimulating factor (GCSF). While our repeated-dose toxicity data suggest that these variants do not elicit any adverse effects or histopathological changes, aggregated GCSF impurity caused sluggishness in animal behavior manifested by a possible muscular injury. Cell assay data revealed that the cys-64-cys74 disulfide bond in reduced GCSF imparts stabilization in absence of the cys-36-cys42 bond. PK data demonstrate variability in half lives of different species when compared to the native GCSF. PD data along with differential expression of JAK-2 and STAT5a genes show that all the tested variants triggered the required signal transduction pathways for neutrophil proliferation and activation.
在质量源于设计的范式下，估计各种与产品相关的变体和杂质对生物治疗剂的安全性和疗效的影响是一个基本要求。鉴于临床研究在这方面的作用有限，我们展示了一种临床前方法来实现这一目标，以粒细胞集落刺激因子 (GCSF) 为例。虽然我们的重复剂量毒性数据表明这些变体不会引起任何不良反应或组织病理学变化，但聚集的 GCSF 杂质导致动物行为迟缓，可能导致肌肉损伤。细胞分析数据显示，还原型 GCSF 中的半胱氨酸 64-半胱氨酸 74 二硫键在没有半胱氨酸 36-半胱氨酸 42 键的情况下赋予其稳定性。PK 数据表明，与天然 GCSF 相比，不同物种的半衰期存在差异。PD 数据以及 JAK-2 和 STAT5a 基因的差异表达表明，所有测试的变体都触发了中性粒细胞增殖和激活所需的信号转导途径。
