Manufacturing processes for polypeptide/protein drugs are designed to ensure robust quality, efficacy and safety. Process differences introduced by follow-on manufacturers may result in changes in quality and clinical outcomes. This study investigated the impact of production methods on the stability and impurities of liraglutide and semaglutide drug substances/products, and the potential impact on drug quality, efficacy and safety.
State-of-the-art analytical methods were used to compare physical and chemical stability, and impurity profiles of drug substances/products from different suppliers. Identified polypeptide-related impurities were evaluated for immunogenicity potential by in silico T cell epitope prediction. Semaglutide immunogenicity in clinical trials (SUSTAIN) was evaluated using a tiered antibody analysis.
Manufacturing scale and process strongly impacted the physical stability of the products. Trace metals increased high-molecular-weight protein formation for liraglutide and semaglutide. Synthetic and recombinant liraglutide produced by five suppliers had distinct impurity profiles compared with the originator. In silico evaluation suggested that new impurities could be immunogenic. Immunogenicity of semaglutide in clinical trials was lower than for liraglutide.
Differences in manufacturing processes affect chemical/physical stability and impurity profile, and may impact immunogenicity. Follow-on versions of liraglutide and semaglutide, and possibly other polypeptides, should be clinically evaluated for efficacy and safety.
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Percentage of bound (B) relative to total (T) radioactivity
Digital single-lens reflex
- EC70–80 :
70–80% maximal effective concentration
Good Manufacturing Practice
Human leukocyte antigen
High-performance liquid chromatography
Inductively Coupled Plasma – Optical Emission Spectroscopy
Liquid chromatography-mass spectrometry
Liraglutide Effect and Action on Diabetes
Limit of quantification
Major histocompatibility complex
Nephelometric turbidity units
Reverse phase high-performance liquid chromatography
Size exclusion chromatography
Semaglutide Unabated Sustainability in Treatment of Type 2 Diabetes
Ultra-performance liquid chromatography
United States Pharmacopeia
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Acknowledgments and Disclosures
The authors would like to thank Bengt-Olof Axelsson, Magnus Bernt, Dorthe Kot Engelund, Frederik Gustafsson, Morten Hach, Mattias Hansson, Karen Hauda, Susanne Hostrup, Heidi Jensen, Magdalena Renee Johansen, Inger Mollerup, Griffin E. Moran, Claus Aagaard Nielsen, Mikkel Due Petersen, Lars Nygaard, David Ørsted, Christian Poulsen, Tina Secher Rasmussen, and Else Marie Agger (Novo Nordisk A/S, Denmark) for review of and input to the manuscript, and to Alexander Jones, PhD, CMPPTM and Sola Neunie, MSc, CMPPTM from AXON Communications (supported by Novo Nordisk) for writing assisstance. The study was sponsored by Novo Nordisk and the SUSTAIN are registered with ClinicalTrials.gov (NCT02054897; NCT01885208; NCT02128932; NCT01930188; NCT02305381; NCT02207374; NCT02254291; NCT01720446).
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Staby, A., Steensgaard, D.B., Haselmann, K.F. et al. Influence of Production Process and Scale on Quality of Polypeptide Drugs: a Case Study on GLP-1 Analogs. Pharm Res 37, 120 (2020). https://doi.org/10.1007/s11095-020-02817-9