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In vitro activity and host impurity detection of GLP-1 analogues
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Zhi-hua ZHENG1, Qian-qian CHEN2, Qian-qian GU1, Pei-qing LIU1, *, Min LI1, *
Acta Pharmaceutica Sinica | 2020, 55(9) : 2145 - 2150
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Acta Pharmaceutica Sinica | 2020, 55(9): 2145-2150
• Original Articles •
In vitro activity and host impurity detection of GLP-1 analogues
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Zhi-hua ZHENG1, Qian-qian CHEN2, Qian-qian GU1, Pei-qing LIU1, *, Min LI1, *
Affiliations
  • 1. School of Pharmaceutical Sciences, National and Local United Engineering Lab of Druggability and New Drug Evaluation, Sun Yat-Sen University, Guangzhou 510006, China
  • 2. Livzon New River Pharmaceutical Co., Ltd., Qingyuan 511515, China
Published: 2020-09-12 doi: 10.16438/j.0513-4870.2020-0307
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Glucagon-like peptide-1 (GLP-1) could increase the level of cyclic adenosine monophosphate (cAMP) in cells to stimulate insulin secretion in β cells of pancreas. So GLP-1 analogues, such as liraglutide, have become new anti-hyperglycemia drugs for type 2 diabetes. In this study, a set of in vitro activity detection method suitable for GLP-1 analogues was established using GLP-1R-GFP (green fluorescent protein, GFP)-HEK293A cells which stably expressing GLP-1 receptor (GLP-1R). After optimizing the detection parameters such as assay sensitivity, cell density, and the incubation condition, the cAMP content level of GLP-1R-GFP-HEK293A cells stimulated by four GLP-1 analogues, such as liraglutide, were detected by homogeneous time-resolved fluorescence (HTRF). The values of concentration for 50% of maximal effect (EC50) of GLP-1 analogues were calculated by cAMP dose-response curve to evaluate the in vitro activity of those drugs. In addition, enzyme-linked immunosorbent assay and quantitative polymerase chain reaction were applied to determine the content of host cell protein and host residual DNA, respectively. This study provides a stable, reliable, and sensitive in vitro activity analysis and host impurity detection method for high-throughput screening of GLP-1 analogues.

glucagon-like peptide-1  /  cyclic adenosine monophosphate  /  homogeneous time-resolved fluorescence  /  concentration for 50% of maximal effect  /  liraglutide
Zhi-hua ZHENG, Qian-qian CHEN, Qian-qian GU, Pei-qing LIU, Min LI. In vitro activity and host impurity detection of GLP-1 analogues[J]. Acta Pharmaceutica Sinica, 2020 , 55 (9) : 2145 -2150 . DOI: 10.16438/j.0513-4870.2020-0307
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Article Info
doi: 10.16438/j.0513-4870.2020-0307
  • Receive Date：2020-03-12
  • Online Date：2026-01-23
  • Published：2020-09-12
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History
  • Received：2020-03-12
  • Revised：2020-04-16
Funding
Affiliations
    1. School of Pharmaceutical Sciences, National and Local United Engineering Lab of Druggability and New Drug Evaluation, Sun Yat-Sen University, Guangzhou 510006, China
    2. Livzon New River Pharmaceutical Co., Ltd., Qingyuan 511515, China
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