2025-02-11 作者：ws 来源：文献：Xenopus GLP-1-inspired discovery of novel GLP-1 receptor agonists as longacting hypoglycemic and insulinotropic agents with significant therapeutic potential
文献链接：https://www.sciencedirect.com/science/article/abs/pii/S0006295217304562?via%3Dihub
作者：Jing Han, Xinyu Chen, Yiyun Wang, Yingying Fei, Feng Zhou, Ying Zhang,
Lin Liu, Pengbin Si, Junjie Fu
原文摘要：
We here report the discovery and therapeutic efficacy of a novel series of glucagon-like peptide-1 (GLP-1) receptor agonists derived from Xenopus GLP-1. First, five amino acid-mutated Xenopus GLP-1s were synthesized, and xGLP-3 with the best acute and long-acting hypoglycemic activity was selected for further modification. Next, PEGylation of xGLP-3 was performed at specific sites, which were determined using cysteine mutagenesis scanning. Twelve PEGylated conjugates tethered with Mal-PEGs of 1, 2, and 5 kDa were synthesized. Conjugates 7b and 7c, which exhibited comparable hypoglycemic and insulinotropic effects to Gly8-GLP-1, were selected for in-depth evaluation. It was found that 7b and 7c exhibited prolonged in vivo half-life and improved pharmacokinetic behaviors. The long-term hypoglycemic effects of 7b and 7c were further confirmed by pre-OGTT and multiple OGTT. Importantly, long-term administration of 7b or 7c in db/db mice achieved beneficial effects on body weight loss, food intake and HbA1c reduction, and glucose tolerance normalization. These preclinical studies indicate the promising role of 7b and 7c as long-acting type 2 diabetes therapeutics. In addition, our research demonstrated the feasibility of developing novel antidiabetic agents based on Xenopus GLP-1.
mPEG - MAL 是一种聚乙二醇（PEG）衍生物。“mPEG” 代表甲氧基聚乙二醇，它是在聚乙二醇链的一端连接了一个甲氧基（-OCH3）。甲氧基的存在使得聚乙二醇链具有一定的化学稳定性，并且这个末端相对比较惰性。“MAL” 代表马来酰亚胺（Maleimide）基团，马来酰亚胺基团连接在聚乙二醇链的另一端。马来酰亚胺基团具有高度的化学反应活性，它可以与含有巯基（-SH）的化合物发生特异性的迈克尔加成反应。基于mPEG - MAL的性能，新型GLP-1受体激动剂合成如下：
图：合成示意
聚乙二醇化XenGLP-1类似物的一般合成路线：
将1 kDa、2 kDa和5 kDa的mPEG-MAL溶解在磷酸钠缓冲液中。然后将多肽加入混合物中，在N2下室温搅拌反应，HPLC监测反应。采用半制备RP所得混合物以一定的流速-HPLC纯化。洗脱液A和B分别由水TFA和甲醇TFA组成，应用条件如下：在一段百分比内的B呈线性梯度。用Thermo LC-MS对纯化的XenGLP-1类似物进行了表征。
图：图谱示意
结论：
该文献成功制备了基于mPEG - MAL合成的新型GLP-1受体激动剂。实验发现，xGLP-3具有最佳的急性和长效降糖活性。在特定位点对xGLP-3进行聚乙二醇化，并通过半胱氨酸诱变扫描进行测定，结果发现，7b和7c的体内半衰期延长，药代动力学行为得到改善。
