Yingxin Wang1, Zhijuan Xia2, Hezhi Xie3, Jiahang Dong4. 1. School of Physical Education, Shandong Normal University, Jinan 250358, Shandong, PR China. 2. Department of Nephrology, The First Affiliated Hospital of University of South China, Hengyang 421001, Hunan, PR China. 3. School of Athletic Training, Guangzhou Sport University, Guangzhou 510000, Guangdong, PR China. Electronic address: txhzhi@outlook.com. 4. School of Physical Education, Shandong Normal University, Jinan 250358, Shandong, PR China. Electronic address: jhang134@outlook.com.
Abstract
AIMS: To design and evaluate the anti-hyperglycemia and overexercise-induced myocardial injury efficacies of a novel long-acting glucagon-like peptide-1 (GLP-1)-based therapeutic peptide in rodent animals. MAIN METHODS: Here, we designed and prepared a new pro-drug, termed RYHSB-1, which was connected by a mutated GLP-1(A8G) and an albumin binding peptide via a protease-cleavable linker. Moreover, isothermal titration calorimetry (ITC) was applied to detect its binding affinity for HSA. GLP-1 release assay was conducted in mouse serum in vitro and quantified using LC-MS/MS method. Modified intraperitoneal glucose tolerance test (IPGTT), chronic efficacies study in rodent animals with overexercise-induced myocardial injury were subjected to evaluate the druggability of RYHSB-1. RESULTS: RYHSB-1 with purity over 99% was prepared and ITC measurement demonstrated high binding affinity for HSA with KD of 0.06 μM. Protease cleavage assay demonstrated slowly controlled-release of transient GLP-1 from RYHSB-1 under the hydrolysis catalyzed by thrombin in vitro. Moreover, IPGTT showed clearly dose-dependent glucose-lowering efficacies of RYHSB-1 within 0.1-0.9 mg/kg. The prolonged anti-diabetic efficacy of RYHSB-1 was further assessed via multiple IPGTTs and hypoglycemic duration test. Furthermore, long-term administration of RYHSB-1 in diabetic mice achieved promising efficacies on hyperglycemia and overexercise-induced myocardial injury. SIGNIFICANCE: RYHSB-1 holds outstanding pharmaceutical potential as an anti- overexercise-induced myocardial injury drug. The strategy of albumin-conjugation also could be applied to other active peptides develop long effecting therapeutic drugs.
AIMS: To design and evaluate the anti-hyperglycemia and overexercise-induced myocardial injury efficacies of a novel long-acting glucagon-like peptide-1 (GLP-1)-based therapeutic peptide in rodent animals. MAIN METHODS: Here, we designed and prepared a new pro-drug, termed RYHSB-1, which was connected by a mutated GLP-1(A8G) and an albumin binding peptide via a protease-cleavable linker. Moreover, isothermal titration calorimetry (ITC) was applied to detect its binding affinity for HSA. GLP-1 release assay was conducted in mouse serum in vitro and quantified using LC-MS/MS method. Modified intraperitoneal glucose tolerance test (IPGTT), chronic efficacies study in rodent animals with overexercise-induced myocardial injury were subjected to evaluate the druggability of RYHSB-1. RESULTS: RYHSB-1 with purity over 99% was prepared and ITC measurement demonstrated high binding affinity for HSA with KD of 0.06 μM. Protease cleavage assay demonstrated slowly controlled-release of transient GLP-1 from RYHSB-1 under the hydrolysis catalyzed by thrombin in vitro. Moreover, IPGTT showed clearly dose-dependent glucose-lowering efficacies of RYHSB-1 within 0.1-0.9 mg/kg. The prolonged anti-diabetic efficacy of RYHSB-1 was further assessed via multiple IPGTTs and hypoglycemic duration test. Furthermore, long-term administration of RYHSB-1 in diabeticmice achieved promising efficacies on hyperglycemia and overexercise-induced myocardial injury. SIGNIFICANCE: RYHSB-1 holds outstanding pharmaceutical potential as an anti- overexercise-induced myocardial injury drug. The strategy of albumin-conjugation also could be applied to other active peptides develop long effecting therapeutic drugs.