Format

Send to

Choose Destination
Eur J Pharm Biopharm. 2016 Nov;108:156-167. doi: 10.1016/j.ejpb.2016.09.003. Epub 2016 Sep 8.

Study of stability and biophysical characterization of ranibizumab and aflibercept.

Author information

1
School of Material Science & Engineering, Nanyang Technological University, Singapore. Electronic address: mmoreno@ntu.edu.sg.
2
School of Material Science & Engineering, Nanyang Technological University, Singapore.
3
School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore.
4
Tan Tock Seng Hospital, Singapore. Electronic address: rupesh_agrawal@ttsh.com.sg.

Abstract

The anti-vascular endothelial growth factor (VEGF) agents such as ranibizumab (Lucentis®) and aflibercept (EyLea®) are currently used as monthly or bimonthly intravitreal injections to treat potentially retinal diseases such as wet age-related macular degeneration (AMD) or diabetic macular edema (DME). Because of the complications associated with repeated intra-vitreal injections, there is considerable interest in developing a sustained delivery system. The purpose of this study was to examine the stability of both therapeutic proteins under physiological conditions as well as when incorporated into drug delivery systems (DDS). First, thermotropic properties in physiological conditions and at different pH values were evaluated by differential scanning calorimetry (DSC) to determine the protein denaturation temperature. Second, the effects of pH and incubation time on conformational changes and aggregation were evaluated by circular dichroism (CD), steady-state tryptophan fluorescence spectroscopy, and size-exclusion chromatography (SEC). Also, the ability of both proteins to bind to VEGF was tested in the aforementioned experimental conditions for up to 30 days. Finally, we investigated the stability of both proteins after a rapid screening method that simulates the first homogenizing step during the protein microencapsulation process. This method allowed the development of stable ranibizumab and aflibercept formulations that may be useful to entrap these proteins into microparticles selecting the most convenient organic solvent and protein stabilizers.

KEYWORDS:

Aggregation state; Anti-vascular endothelial growth factor (VEGF) proteins; Biophysical characterization; Diabetic macular edema (DME); Emulsion; Excipients; Stability; Wet age related macular degeneration (AMD)

PMID:
27615995
DOI:
10.1016/j.ejpb.2016.09.003
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center