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Comparative Stability Studies of Different Infliximab and Biosimilar CT-P13 Clinical Solutions by Combined Use of Physicochemical Analytical Techniques and Enzyme-Linked Immunosorbent Assay (ELISA)

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Abstract

Background

There are two products in which infliximab is the active pharmaceutical ingredient. These are Remicade® (INF; reference product) and Remsima™/Inflectra™ (CT-P13; infliximab biosimilar). Remsima™/Inflectra™ are bioidentical products. Different recommendations have been made for the clinical solutions of each brand (Remicade® or Remsima™/Inflectra™) despite the manufacturer of the biosimilar claiming high levels of similarity to the innovator.

Objective

The objective of this study was to assess and compare stability against degradation and over time of different clinical infliximab solutions prepared from Remicade® and from Remsima™/Inflectra™ using a suitable set of characterization methods in line with the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) recommendations.

Methods

Reconstituted solutions of INF and CT-P13 and dilutions as used in hospital were stored in glass vials (10 and 2 mg/mL) or in polyolefin infusion bags (0.4 mg/mL) refrigerated between 2 and 8 °C for 2 weeks. Regarding the physicochemical properties, the distribution of the particulates were studied over a range of 0.001–1 µm by dynamic light scattering (DLS) and oligomers up to 8 monomer were analyzed by native size-exclusion ultra-high-performance liquid chromatography with ultraviolet (UV)-visible detection coupled to (native) mass spectrometry (SE/UHPLC-UV-(native) MS); mass spectrometry was also used to evaluate natural aggregates and isoform profile; DLS was also employed to detect gross conformational changes by tracking the hydrodynamic radius (HR). The secondary structure of the proteins was studied by far UV circular dichroism (CD). The tertiary structure was investigated by intrinsic tryptophan fluorescence (IT-F). Reverse-phase ultra-high-performance liquid chromatography with UV detection (RP/UHPLC-UV) was used to analyze intact INF and CT-P13 for quantification purposes. Functionality was evaluated via the biological activity measured by the extension of the immunological reaction of the INF and the CT-P13 with its antigen, i.e., the tumor necrosis factor-α by enzyme-linked immunosorbent assay (ELISA).

Results

The stress applied to INF and CT-P13 solutions showed similar levels of aggregate formation, structural variation, and chemical modifications. The only noteworthy difference between INF and CT-P13 was detected in their behavior to freeze–thaw cycles, in which CT-P13 showed slightly more robustness. INF and CT-P13 showed identical CD spectra, similar to those reported for IgG1 in which there is dominance in β sheet secondary structures; this typical conformation remained unmodified over time in INF and CT-P13. No significant changes were detected in the tertiary structure and no aggregates process was noticed over the time studied. Polydispersity slightly increased for the most concentrated solutions, while there were no meaningful differences in the HR in the solutions over time. The concentration of INF and CT-P13 also remained constant. Differences in the native isoform MS profile were detected, as expected by the different glycosylation pattern, with no important modification over time. Functionality was maintained over the test period (60 days) and was similar in all the solutions tested, with no differences between INF and biosimilar solutions.

Conclusions

High levels of similarity were noticed in the behavior of INF and CT-P13 when subjected to stress. When stored refrigerated at between 2 and 8 °C and prepared as normally used in the hospital pharmacy, all solutions showed physicochemical and functional stability for all the concentrations tested and all containers, at least for the 14-day test period.

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References

  1. Committee for Medicinal Products for Human Use. Remsima assessment report EMA/CHMP/589317/2013. 2013. https://www.ema.europa.eu/documents/assessment-report/remsima-epar-public-assessment-report_en.pdf. Accessed 20 Nov 2018.

  2. Committee for Medicinal Products for Human Use. Inflectra assessment report EMA/CHMP/589422/2013. 2013. https://www.ema.europa.eu/documents/assessment-report/inflectra-epar-public-assessment-report_en.pdf. Accessed 20 Nov 2018.

  3. Wong M, Ziring D, Korin Y, Desai S, Kim S, Lin J, et al. TNFα blockade in human diseases: mechanisms and future directions. Clin Immunol. 2008;126:121–36.

    Article  CAS  PubMed  Google Scholar 

  4. Couriel DR, Hicks K, Giralt S, Champlin RE. Role of tumor necrosis factor-alpha inhibition with inflixiMAB in cancer therapy and hematopoietic stem cell transplantation. Curr Opin Oncol. 2000;12:582–7.

    Article  CAS  PubMed  Google Scholar 

  5. Blair HA, Deeks ED. Infliximab biosimilar (CT-P13; infliximab-dyyb): a review in autoimmune inflammatory diseases. BioDrugs. 2016;30:469–80.

    Article  CAS  PubMed  Google Scholar 

  6. Pisupati K, Benet A, Tian Y, Okbazghi S, Kang J, Ford M, et al. Biosimilarity under stress: a forced degradation study of Remicade® and Remsima™. MAbs. 2017;9:1197–209.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Young BL, Khan MA, Chapman TJ, Parry R, Connolly MA, Watts AG. Evaluation of the physicochemical and functional stability of diluted REMSIMA® upon extended storage—a study compliant with NHS (UK) guidance. Int J Pharm. 2015;496:421–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Vieillard V, Astier A, Sauzay C, Paul M. One-month stability study of a biosimilar of infliximab (Remsima®) after dilution and storage at 4 °C and 25 °C. Ann Pharm Fr. 2017;75:17–29.

    Article  CAS  PubMed  Google Scholar 

  9. Tokhadze N, Chennell P, Le Basle Y, Sautou V. Stability of infliximab solutions in different temperature and dilution conditions. J Pharm Biomed Anal. 2018;150:386–95.

    Article  CAS  PubMed  Google Scholar 

  10. EMA. Remicade®, annex 1: summary of product characteristics. 2018. https://www.ema.europa.eu/documents/product-information/remicade-epar-product-information_en.pdf. Accessed 20 Nov 2018.

  11. EMA. Remicade®, annex 1: summary of product characteristics. 1999. https://ec.europa.eu/health/documents/community-register/1999/199908133334/anx_3334_en.pdf. Accessed 20 Nov 2018.

  12. Alsaddique JA, Pabari RM, Ramtoola Z. Effect of thermal and shear stressors on the physical properties, structural integrity and biological activity of the anti-TNF-alpha monoclonal antibody, infliximab. Curr Pharm Biotechnol. 2016;17:905–14.

    Article  CAS  PubMed  Google Scholar 

  13. Ikeda R, Vermeulen LC, Lau E, Jiang Z, Saha S, Reichelderfer M, et al. Stability of infliximab in polyvinyl chloride bags. Am J Heal Pharm. 2012;69:1509–12.

    Article  CAS  Google Scholar 

  14. EMA. Remsima®, annex 1: summary of product characteristics. 2010. https://www.ema.europa.eu/documents/product-information/remsima-epar-product-information_en.pdf. Accessed 06 Feb 2019.

  15. EMA. Inflectra®, annex 1: summary of product characteristics. 2010. https://www.ema.europa.eu/documents/product-information/inflectra-epar-product-information_en.pdf. Accessed 06 Feb 2019.

  16. ICH Expert Working Group. Specifications: test procedures and acceptance criteria for biotechnological/biological products Q6B. 1999. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q6B/Step4/Q6B_Guideline.pdf. Accessed 27 Nov 2018.

  17. Visser J, Feuerstein I, Stangler T, Schmiederer T, Fritsch C, Schiestl M. Physicochemical and functional comparability between the proposed biosimilar rituximab GP2013 and originator rituximab. BioDrugs. 2013;27:495–507.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Martínez-Ortega A, Herrera A, Salmerón-García A, Cabeza J, Cuadros-Rodríguez L, Navas N. Validated reverse phase HPLC diode array method for the quantification of intact bevacizumab, infliximab and trastuzumab for long-term stability study. Int J Biol Macromol. 2018;116:993–1003.

    Article  CAS  PubMed  Google Scholar 

  19. Hernández-Jiménez J, Martínez-Ortega A, Salmerón-García A, Cabeza J, Prados JC, Ortíz R, et al. Study of aggregation in therapeutic monoclonal antibodies subjected to stress and long-term stability tests by analyzing size exclusion liquid chromatographic profiles. Int J Biol Macromol. 2018;118:511–24.

    Article  CAS  PubMed  Google Scholar 

  20. ICH Expert Working Group. Stability testing: photostability testing of new drug substances and products. Q1B. 1996. https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q1B/Step4/Q1B_Guideline.pdf. Accessed 27 Nov 2018.

  21. Pérez-Robles R, Cuadros-Rodríguez L, Salmerón-García A, Navas N. Development and validation of a (RP)UHPLC-UV-(HESI/Orbitrap)MS method for the identification and quantification of mixtures of intact therapeutical monoclonal antibodies using a monolithic column. J Pharm Biomed Anal. 2018;159:437–48.

    Article  CAS  PubMed  Google Scholar 

  22. Suárez I. ELISA methodology to study the stability of biotechnological medicines [in Spanish]. University of Granada; 2016. http://hdl.handle.net/10481/47150. Accessed 27 Nov 2018.

  23. Suárez I, Salmerón-García A, Cabeza J, Capitán-Vallvey LF, Navas N. Development and use of specific ELISA methods for quantifying the biological activity of bevacizumab, cetuximab and trastuzumab in stability studies. J Chromatogr B Anal Technol Biomed Life Sci. 2016;1032:155–64.

    Article  CAS  Google Scholar 

  24. Wang Y, Fei D, Vanderlaan M, Song A. Biological activity of bevacizumab, a humanized anti-VEGF antibody in vitro. Angiogenesis. 2004;7:335–45.

    Article  CAS  PubMed  Google Scholar 

  25. Vermeer AWP, Bremer MGEG, Norde W. Structural changes of IgG induced by heat treatment and by adsorption onto a hydrophobic Teflon surface studied by circular dichroism spectroscopy. Biochim Biophys Acta. 1998;1425:1–12.

    Article  CAS  PubMed  Google Scholar 

  26. Pabari RM, Ryan B, McCarthy C, Ramtoola Z. Effect of microencapsulation shear stress on the structural integrity and biological activity of a model monoclonal antibody, trastuzumab. Pharmaceutics. 2011;3:510–24.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Langridge TD, Tarver MJ, Whitten ST. Temperature effects on the hydrodynamic radius of the intrinsically disordered N-terminal region of the p53 protein. Proteins Struct Funct Bioinform. 2014;82:668–78.

    Article  CAS  Google Scholar 

  28. Hernández-Jiménez J, Salmerón-García A, Cabeza J, Vélez C, Capitán-Vallvey LF, Navas N. The effects of light-accelerated degradation on the aggregation of marketed therapeutic monoclonal antibodies evaluated by size-exclusion chromatography with diode array detection. J Pharm Sci. 2016;105:1405–18.

    Article  CAS  PubMed  Google Scholar 

  29. Lee C, Jeong M, Lee JAJ, Seo S, Cho SC, Zhang W, et al. Glycosylation profile and biological activity of Remicade® compared with Flixabi® and Remsima®. MAbs. 2017;9:968–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors would like to thank the Hospital Pharmacy Unit of the University Hospital San Cecilio (Grenada, Spain) for kindly supplying all the medicine samples and the Biomedical Research Foundation “Alejandro Otero” (FIBAO) for the support given during the course of this research.

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Authors and Affiliations

  1. Department of Analytical Chemistry, Sciences Faculty/Biomedical Research Institute ibs.Granada, University of Granada, Fuentenueva Avenue s/n, 18071, Granada, Spain

    Jesús Hermosilla, Ricardo Sánchez-Martín, Raquel Pérez-Robles, Luis Cuadros-Rodríguez & Natalia Navas

  2. UGC Farmacia Hospitalaria, Biomedical Research Institute ibs.Granada, Hospital Universitario San Cecilio de Granada, 18012, Granada, Spain

    Antonio Salmerón-García & Jose Cabeza

  3. Department of Physical Chemistry, Faculty of Sciences, University of Granada, 18071, Granada, Spain

    Salvador Casares

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  1. Jesús Hermosilla
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Correspondence to Natalia Navas.

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Conflict of interest

Jesús Hermosilla, Ricardo Sánchez Martín, Raquel Pérez-Robles, Antonio Salmerón-García, Salvador Casares, Jose Cabeza, Luis Cuadros-Rodríguez, and Natalia Navas declare that they have no conflict of interest related to the contents of this article that might influence the results of the investigation.

Funding

This study was entirely funded by Project FIS: PI-17/00547 (Instituto Carlos III, Ministerio de Economía y Competitividad, Spain), which means that it was also partially supported by European Regional Development Funds (ERDF).

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Hermosilla, J., Sánchez-Martín, R., Pérez-Robles, R. et al. Comparative Stability Studies of Different Infliximab and Biosimilar CT-P13 Clinical Solutions by Combined Use of Physicochemical Analytical Techniques and Enzyme-Linked Immunosorbent Assay (ELISA). BioDrugs 33, 193–205 (2019). https://doi.org/10.1007/s40259-019-00342-9

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  • DOI: https://doi.org/10.1007/s40259-019-00342-9

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