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Influence of Production Process and Scale on Quality of Polypeptide Drugs: a Case Study on GLP-1 Analogs

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Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

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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Abbreviations

ADA:

Anti-drug antibody

AU:

Arbitrary units

B/T:

Percentage of bound (B) relative to total (T) radioactivity

DP:

Drug product

DPP-IV:

Dipeptidyl peptidase-IV

DS:

Drug substance

dSLR:

Digital single-lens reflex

EC70–80 :

70–80% maximal effective concentration

GLP-1:

Glucagon-like peptide-1

GMP:

Good Manufacturing Practice

HLA:

Human leukocyte antigen

HMWP:

High-molecular-weight protein

HPLC:

High-performance liquid chromatography

ICP–OES:

Inductively Coupled Plasma – Optical Emission Spectroscopy

LC-MS:

Liquid chromatography-mass spectrometry

LEAD:

Liraglutide Effect and Action on Diabetes

LOQ:

Limit of quantification

MHC:

Major histocompatibility complex

MS:

Mass spectrometry

nAb:

Neutralizing antibody

NTU:

Nephelometric turbidity units

PEG:

Polyethylene glycol

RP-HPLC:

Reverse phase high-performance liquid chromatography

s.c.:

Subcutaneous

SEC:

Size exclusion chromatography

SUSTAIN:

Semaglutide Unabated Sustainability in Treatment of Type 2 Diabetes

ThT:

Thioflavin T

UPLC:

Ultra-performance liquid chromatography

USP:

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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AS contributed to study conception and design, data analysis, and the writing and critical revision of the manuscript at all stages of development. DBS contributed to study conception and design, data analysis, and the writing and critical revision of the manuscript at all stages of development. CB contributed to study conception and design, data collection, data analysis, and the writing and critical revision of the manuscript at all stages of development. DA contributed to study conception and design, and critical revision of the manuscript at all stages of development. HBT contributed to writing and critical revision of the manuscript at all stages of development. JSM contributed to study conception and design, data analysis, and the writing and critical revision of the manuscript at all stages of development. KFH contributed to study conception and design, material preparation, data collection and analysis, and the writing and critical revision of the manuscript at all stages of development. LTS contributed to study conception and design, and critical revision of the manuscript at all stages of development. NV contributed to data collection and analysis, and the writing and critical revision of the manuscript at all stages of development. OS contributed to study conception and design, data analysis, and the writing and critical revision of the manuscript at all stages of development. All authors approved the final version.

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Correspondence to Arne Staby.

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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

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