Artificial sunlight; Dilution effect; Mechanical stability; Monoclonal antibody; Photostability; Protein drug product; Antibodies, Monoclonal; Antineoplastic Agents, Immunological; Excipients; Ipilimumab; Antibodies, Monoclonal/chemistry; Antibodies, Monoclonal/pharmacology; Excipients/chemistry; Humans; Protein Stability; Drug Stability; Biotechnology; Pharmaceutical Science; General Medicine
Abstract :
[en] The stability of the monoclonal antibody Ipilimumab, the active ingredient of Yervoy®, used for the treatment of different types of cancer, has been investigated. Shaking/temperature, light exposure and dilution, protein drug renowned stressors, were applied on a 30-45-day series of experiments to observe the physicochemical and biological behavior of the molecule. Ipilimumab demonstrated stability under shaking and heat up to 45 days, without any unfolding during the induced combined stressors. Under artificial sunlight, the mAb showed to be sensitive even under the minimum dose tested (720 kJ/m2) with formation of aggregates, particularly when diluted in glucose solution. The light-induced soluble aggregates were higher in the case of diluted samples irradiated with much higher light doses (10460 kJ/m2). The aggregation of Ipilimumab took place also by irradiating the non-diluted formulation, indicating that the excipients did not protect completely the drug from photodegradation. Amino acid oxidation and deamidation were found. Anyway, after irradiation with both light doses, soluble Ipilimumab maintained its typical β-sheets structure, and the tertiary structure was nearly maintained compared to the dark. As an additional stressor test, the effect of dilution on the formulation was monitored by using a saline solution (1 mg/mL Ipilimumab) applied during hospital infusion. After two days from dilution, the protein exhibited aggregation and chemical modifications including oxidation and deamidation. When stability conditions were compromised, the viability of human cell lines treated with the stressed formulation slight decreased suggesting low potential biological toxicity of the modified mAb. As this study has demonstrated the susceptibility of Ipilimumab to light, specific solutions, and excipients as well as the use of safe light in manufacturing, handling, and storage of this drug should be promoted. Moreover, the use of proper primary and secondary packaging should be indicated to avoid the detrimental effect of light on the mAb structure and efficacy. A detailed understanding of Ipilimumab physicochemical properties, integrity, and stability could assure the best storage and manipulation conditions for its safe and successful application in cancer therapy.
Fongaro, Benedetta ; Université de Liège - ULiège > Département des sciences de la vie > Centre d'Ingénierie des Protéines (CIP) ; Department of Pharmaceutical and Pharmacological Sciences, Via Marzolo, 5, 35131 Padova, Italy
Cian, Valentina; Department of Pharmaceutical and Pharmacological Sciences, Via Marzolo, 5, 35131 Padova, Italy
Gabaldo, Francesca; Department of Pharmaceutical and Pharmacological Sciences, Via Marzolo, 5, 35131 Padova, Italy
De Paoli, Giorgia; Molecular and Clinical Medicine, School of Medicine, University of Dundee Nethergate, Dundee, Scotland DD1 4HN, UK
Miolo, Giorgia ; Department of Pharmaceutical and Pharmacological Sciences, Via Marzolo, 5, 35131 Padova, Italy. Electronic address: giorgia.miolo@unipd.it
Polverino de Laureto, Patrizia; Department of Pharmaceutical and Pharmacological Sciences, Via Marzolo, 5, 35131 Padova, Italy. Electronic address: patrizia.polverinodelaureto@unipd.it
Language :
English
Title :
Managing antibody stability: Effects of stressors on Ipilimumab from the commercial formulation to diluted solutions.
Publication date :
July 2022
Journal title :
European Journal of Pharmaceutics and Biopharmaceutics
H2020 - 101007939 - RealHOPE - Real World Handling of Protein Drugs - Exploration, Evaluation and Education
Funders :
IMI - Innovative Medicines Initiative [BE] EU - European Union [BE]
Funding text :
This work was supported by IMI Project: Real World Handling of Protein Drugs-Exploration, Evaluation and Education - RealHOPE No. 101007939. We thank Dr. Marino Bellini for the technical assistance in mass spectrometry.
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