College of Animal Sciences, Zhejiang University, Hangzhou 310058, PR China; Hangzhou RunChongGuiMei Bio-tech Co., Ltd., Xiao Shan, Hangzhou 310058, PR China.
National Research Center for Veterinary Medicine, Road Cuiwei, High-Tech District, Luoyang 471003, PR China.
Life Sci. 2020 Jul 15;253:117651. doi: 10.1016/j.lfs.2020.117651. Epub 2020 Apr 15.
To investigate the combination of dimerization and PEGylation to enhance the receptor activation and in vivo stability of Oxyntomodulin (OXM).
All LDM peptides were produced by using standard method of solid phase synthesis. The in vitro effects of LDM peptides were assessed by glucagon-like peptide-1 receptor (GLP-1R) and glucagon receptor (GcgR) binding test and Proteolytic stability test. Subsequently, saline, Liraglutide and three doses of LDM-3 treated groups were subjected to the evaluation of aute and long-term efficacy.
Five long-acting OXM conjugates, termed LDM-1 to LDM-5, were designed using cysteine (Cys)-specific modification reaction including the activated PEG, bisMal-NH, and OXM-Cys, and all prepared with high purity. LDM-3 exhibited greater GLP-1R and GcgR activation and ameliorative serum stability. In addition, LDM-3 was identified with enhanced insulinotropic and glycemic abilities in the gene knockout mice. The prolonged glucose-lowering effects of the LDM-3 were proved by hypoglycemic duration test and multiple oral glucose tolerance tests (OGTTs) in the diet-induced obesity (DIO) mice. Furthermore, the pharmacokinetic tests in Sprague Dawley (SD) rat and cynomolgus monkey exhibited the lifespans of LDM-3 at 90 nmol·kg were 101.5 h and 119.4 h, respectively. Nevertheless, consecutive 8-week administration of LDM-3 improved the cumulative body weight gain, food intake, % HbA1c, glucose tolerance and the pancreatic of the obese mice.
LDM-3, as a dual GLP-1R and GcgR agonist, holds potential to be developed as a promising therapeutic candidate for both diabetes and obesity.
研究二聚化和聚乙二醇化的结合如何增强胰高血糖素样肽-1 受体（GLP-1R）和胰高血糖素受体（GcgR）的激活以及胰高血糖素样肽-1（OXM）的体内稳定性。
所有 LDM 肽均通过固相合成的标准方法生产。通过 GLP-1R 和 GcgR 结合试验和蛋白水解稳定性试验评估 LDM 肽的体外作用。随后，对盐水、利拉鲁肽和三个剂量的 LDM-3 治疗组进行急性和长期疗效评估。
设计了五种长效 OXM 缀合物，分别命名为 LDM-1 至 LDM-5，使用半胱氨酸（Cys）特异性修饰反应，包括激活的 PEG、双 Mal-NH 和 OXM-Cys，所有这些都具有很高的纯度。LDM-3 表现出更大的 GLP-1R 和 GcgR 激活作用和改善的血清稳定性。此外，LDM-3 在基因敲除小鼠中被鉴定出具有增强的胰岛素分泌和降血糖能力。在饮食诱导肥胖（DIO）小鼠中，通过低血糖持续时间试验和多次口服葡萄糖耐量试验（OGTT）证实了 LDM-3 的延长降血糖作用。此外，在 Sprague Dawley（SD）大鼠和食蟹猴中的药代动力学试验表明，LDM-3 的半衰期在 90 nmol·kg 时分别为 101.5 h 和 119.4 h。然而，连续 8 周给予 LDM-3 可改善肥胖小鼠的累积体重增加、食物摄入量、%HbA1c、葡萄糖耐量和胰腺。
LDM-3 作为一种双重 GLP-1R 和 GcgR 激动剂，有潜力开发成为治疗糖尿病和肥胖症的有前途的候选药物。
