Stability study over time of clinical solutions of ziv-aflibercept prepared in infusion bags using a proper combination of physicochemical and functional strategies

Jesús Hermosilla; Raquel Pérez-Robles; Antonio Salmerón-García; Salvador Casares; Jose Cabeza; Natalia Navas

doi:10.1016/j.jpba.2021.114209

Stability study over time of clinical solutions of ziv-aflibercept prepared in infusion bags using a proper combination of physicochemical and functional strategies

J Pharm Biomed Anal. 2021 Sep 5:203:114209. doi: 10.1016/j.jpba.2021.114209. Epub 2021 Jun 16.

Authors

Jesús Hermosilla¹, Raquel Pérez-Robles¹, Antonio Salmerón-García², Salvador Casares³, Jose Cabeza², Natalia Navas⁴

Affiliations

¹ Department of Analytical Chemistry, Sciences Faculty, Instituto de Investigación Biosanitaria (ibs.Granada), University of Granada, E-18071, Granada, Spain.
² Department of Clinical Pharmacy, Instituto de Investigación Biosanitaria (ibs.Granada), San Cecilio University Hospital, 18016, Granada, Spain.
³ Department of Physical Chemistry, Sciences Faculty, University of Granada, E-18071, Granada, Spain.
⁴ Department of Analytical Chemistry, Sciences Faculty, Instituto de Investigación Biosanitaria (ibs.Granada), University of Granada, E-18071, Granada, Spain. Electronic address: natalia@ugr.es.

PMID: 34153938
DOI: 10.1016/j.jpba.2021.114209

Abstract

A range of biopharmaceutical products are used to target Vascular Endothelial Growth Factor (VEGF), including Eylea® (aflibercept, AFL) and Zaltrap® (ziv-aflibercept, ziv-AFL). The first is indicated for ophthalmological diseases such as neovascular (wet) age-related macular degeneration, while the second is used in the treatment of metastatic colorectal cancer. The stability of AFL in prefilled syringes has been widely studied; however, no research has yet been done on the stability of ziv-AFL in polyolefin infusion bags. Therefore, the purpose of the present research is to evaluate the stability of ziv-AFL (Zaltrap®) clinical solutions prepared under aseptic conditions in polyolefin infusion bags at two different concentrations, i.e. 4.0 and 0.6 mg/mL, and stored refrigerated in darkness at 2-8 °C for 14 days. With that aim, the ziv-AFL clinical solutions were assessed by analysing changes in its physicochemical and functional properties. The distribution of the particulates was studied over a range of 0.001-10 μm by Dynamic Light Scattering (DLS); oligomers were analysed by Size-Exclusion High-Performance Chromatography with Diode Array Detection (SE/HLPC-DAD); the secondary structure of the protein was studied by far UV Circular Dichroism (CD) and the tertiary structure by Intrinsic Tryptophan Fluorescence (IT-F) and Intrinsic Protein Fluorescence (IP-F); charge variants were assessed by Strong Cation Exchange Ultra-High-Performance Chromatography with UV detection (SCX/UHPLC-UV); functionality was evaluated by ELISA by measuring the biological activity as manifested in the extension of the immunological reaction of the ziv-AFL with its antigen (VEGF). Neither aggregation nor oligomerization were detected by the techniques mentioned above. Secondary and tertiary structures remained unchanged over the 14-day period, as did charge variants. The functionality observed initially was maintained along time. Therefore, it could be proposed that the ziv-AFL clinical solutions studied showed great physicochemical and functional stability over a period of two weeks, regardless of the concentration, i.e. 4 or 0.6 mg/mL.

Keywords: Fc-fusion protein; Infusion bags; Stability study; Ziv-Aflibercept.

MeSH terms

Infusions, Parenteral
Receptors, Vascular Endothelial Growth Factor*
Recombinant Fusion Proteins
Vascular Endothelial Growth Factor A*

Substances

Recombinant Fusion Proteins
Vascular Endothelial Growth Factor A
aflibercept
Receptors, Vascular Endothelial Growth Factor