Engineering an enhanced thrombin-based GLP-1 analog with long-lasting glucose-lowering and efficient weight reduction,RSC Advances

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Engineering an enhanced thrombin-based GLP-1 analog with long-lasting glucose-lowering and efficient weight reduction
RSC Advances ( IF 3.9 ) Pub Date : 2019-09-27 , DOI: 10.1039/c9ra06771j
Hongchao Pan ₁ , Yini Xie ₂ , Wenying Lu ₃ , Yin Chen ₄ , Zhao Lu ₅ , Jun Zhen ₅ , Weiwei Wang ₃ , Anquan Shang ₆

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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 in vivo 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. In vitro 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 vitro 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⁻¹. 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⁻¹) 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.

中文翻译：

设计一种增强型基于凝血酶的 GLP-1 类似物，具有长效降糖和有效减轻体重的作用

肽被认为是有效的治疗药物，主要是由于其对靶标的精细效力和选择性。然而，肽类药物的开发和临床应用受到体内较差的严重限制。寿命。在这里，我们设计了一种改进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。使用胰高血糖素样肽-1 (GLP-1) 作为模型，两个新的长效 GLP-1 类似物（称为 XTS1 和 XTS2）包含一个白蛋白结合结构域、一个蛋白酶可切割的接头和一个突变的 GLP-1（ A8Aib) 旨在证明 GLP-1 由于血浆凝血酶 (TBN) 消化而持续释放。制备了两种 XTS 肽，纯度高 (>99%)，分子量准确，分别通过反向高效液相色谱法和质谱法测定。体外表面等离子共振的测量表明，与 XTS2 相比，XTS1 与所有物种的血清白蛋白具有更高的亲和力。XTS1在人血浆中的体外代谢稳定性也优于XTS2。XTS 肽的蛋白酶切割测定结果表明，在凝血酶催化水解后，瞬时 GLP-1 从 XTS1 和 XTS2 混合物中受控释放。然后腹腔葡萄糖耐量试验（IPGTT）表明XTS1的降糖作用在0.1-0.9 mg kg ^{-1范围内呈剂量依赖性。}. 此外，在多重 IPGTT 和降血糖持续时间测试中，与利拉鲁肽相比，XTS1 显示出相似的降血糖强度和显着更长的作用持续时间。分别在大鼠和食蟹猴中单次皮下施用 XTS1 (0.9 mg kg ^{-1 ) 后观察到明显延长的～2.3 和～3.5 天的血浆半衰期。}此外，在 db/db 小鼠中每周两次皮下注射 XTS1 对体重、血红蛋白 A1C (HbA1C) 降低和胰腺 β 细胞功能具有长期的有益作用。这些研究支持 XTS1 作为治疗 T2DM 的治疗药物发挥潜力。

更新日期：2019-09-27

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