Pan Hongchao, Xie Yini, Lu Wenying, Chen Yin, Lu Zhao, Zhen Jun, Wang Weiwei, Shang Anquan
Department of Laboratory Medicine, Shanghai Simple Gene Medical Laboratory Shanghai 200025 P.R. China.
Department of Laboratory Medicine, The People's Hospital of Jiedong Jieyang 515500 P. R. China.
RSC Adv. 2019 Sep 27;9(53):30707-30714. doi: 10.1039/c9ra06771j. eCollection 2019 Sep 26.
Peptides are considered as potent therapeutic drugs primarily due to the exquisite potency and selectivity to targets. However, the development and clinical application of peptide drugs were severely limited by the poor lifespans. Here, we designed an improved small albumin-binding polypeptide that can associate with human serum albumin (HSA) and liberate the bioactive peptide. Using glucagon-like peptide-1 (GLP-1) as a model, two new long-lasting GLP-1 analogs (termed XTS1 and XTS2) containing an albumin-binding domain, a protease-cleavable linker and a mutated GLP-1(A8Aib) were designed to demonstrate the sustained release of GLP-1 due to the plasma thrombin (TBN) digestion. Two XTS peptides were prepared of high purity (>99%) and accurate molecular weight determined by reversed high-performance liquid chromatography and mass spectrometry, respectively. measurements of surface plasmon resonance indicated that XTS1 associate with serum albumins of all species with higher affinity compared with XTS2. Metabolic stability of XTS1 in human plasma was also better than that of XTS2. Protease cleavage assay results of XTS peptides demonstrated the controlled-release of transient GLP-1 from the XTS1 and XTS2 mixture after thrombin-catalyzed hydrolysis. Then the intraperitoneal glucose tolerance test (IPGTT) showed that the glucose-lowering efficacies of XTS1 were in a dosage-dependent manner within the range of 0.1-0.9 mg kg. In addition, XTS1 showed similar hypoglycemic intensity and significantly longer action duration compared to Liraglutide in both multiple IPGTTs and hypoglycemic duration test. Apparently extended plasma half-lives of ∼2.3 and ∼3.5 days were observed after a single subcutaneous administration of XTS1 (0.9 mg kg) in rats and cynomolgus monkeys, respectively. Furthermore, twice-weekly subcutaneously dosed XTS1 in db/db mice achieved long-term beneficial effects on body weight, hemoglobin A1C (HbA1C) lowering and the function of pancreatic beta cells. These studies support that XTS1 exerts potential as a therapeutic drug for the treatment of T2DM.
肽类主要因其对靶点的高度效力和选择性而被视为有效的治疗药物。然而，肽类药物的开发和临床应用受到其较短半衰期的严重限制。在此，我们设计了一种改良的小白蛋白结合多肽，它可以与人血清白蛋白（HSA）结合并释放生物活性肽。以胰高血糖素样肽-1（GLP-1）为模型，设计了两种新的长效GLP-1类似物（称为XTS1和XTS2），它们含有白蛋白结合结构域、蛋白酶可裂解的连接子和突变的GLP-1（A8Aib），以证明由于血浆凝血酶（TBN）消化导致GLP-1的持续释放。分别通过反相高效液相色谱和质谱制备了两种高纯度（>99%）且分子量准确的XTS肽。表面等离子体共振测量表明，与XTS2相比，XTS1与所有物种的血清白蛋白结合亲和力更高。XTS1在人血浆中的代谢稳定性也优于XTS2。XTS肽的蛋白酶切割试验结果表明，凝血酶催化水解后，XTS1和XTS2混合物中短暂的GLP-1可实现控释。然后，腹腔内葡萄糖耐量试验（IPGTT）表明，在0.1 - 0.9 mg/kg范围内，XTS1的降糖效果呈剂量依赖性。此外，在多次IPGTT和降糖持续时间试验中，与利拉鲁肽相比，XTS1显示出相似的降糖强度和显著更长的作用持续时间。在大鼠和食蟹猴中单次皮下注射XTS1（0.9 mg/kg）后，分别观察到血浆半衰期明显延长至约2.3天和约3.5天。此外，每周两次皮下给药XTS1于db/db小鼠，对体重、糖化血红蛋白A1C（HbA1C）降低和胰腺β细胞功能产生长期有益影响。这些研究支持XTS1作为治疗2型糖尿病的治疗药物具有潜力。
