Characterization, Stability, and In Vivo Efficacy Studies of Recombinant Human CNTF and Its Permeation into the Neural Retina in Ex Vivo Organotypic Retinal Explant Culture Models

Show full item record



Permalink

http://hdl.handle.net/10138/318749

Citation

Itkonen , J , Annala , A , Tavakoli , S , Arango-Gonzalez , B , Ueffing , M , Toropainen , E , Ruponen , M , Casteleijn , M G & Urtti , A 2020 , ' Characterization, Stability, and In Vivo Efficacy Studies of Recombinant Human CNTF and Its Permeation into the Neural Retina in Ex Vivo Organotypic Retinal Explant Culture Models ' , Pharmaceutics , vol. 12 , no. 7 , 611 . https://doi.org/10.3390/pharmaceutics12070611

Title: Characterization, Stability, and In Vivo Efficacy Studies of Recombinant Human CNTF and Its Permeation into the Neural Retina in Ex Vivo Organotypic Retinal Explant Culture Models
Author: Itkonen, Jaakko; Annala, Ada; Tavakoli, Shirin; Arango-Gonzalez, Blanca; Ueffing, Marius; Toropainen, Elisa; Ruponen, Marika; Casteleijn, Marco G.; Urtti, Arto
Contributor: University of Helsinki, Division of Pharmaceutical Biosciences
University of Helsinki, Drug Delivery Unit
University of Helsinki, University of Eastern Finland
University of Helsinki, Division of Pharmaceutical Biosciences
University of Helsinki, Drug Research Program
Date: 2020-07
Language: eng
Number of pages: 30
Belongs to series: Pharmaceutics
ISSN: 1999-4923
URI: http://hdl.handle.net/10138/318749
Abstract: Ciliary neurotrophic factor (CNTF) is one of the most studied neuroprotective agents with acknowledged potential in treating diseases of the posterior eye segment. Although its efficacy and mechanisms of action in the retina have been studied extensively, it is still not comprehensively understood which retinal cells mediate the therapeutic effects of CNTF. As with therapeutic proteins in general, it is poorly elucidated whether exogenous CNTF administered into the vitreous can enter and distribute into the retina and hence reach potentially responsive target cells. Here, we have characterized our purified recombinant human CNTF (rhCNTF), studied the protein's in vitro bioactivity in a cell-based assay, and evaluated the thermodynamic and oligomeric status of the protein during storage. Biological activity of rhCNTF was further evaluated in vivo in an animal model of retinal degeneration. The retinal penetration and distribution of rhCNTF after 24 h was studied utilizing two ex vivo retina models. Based on our characterization findings, our rhCNTF is correctly folded and biologically active. Moreover, based on initial screening and subsequent follow-up, we identified two buffers in which rhCNTF retains its stability during storage. Whereas rhCNTF did not show photoreceptor preservative effect or improve the function of photoreceptors in vivo, this could possibly be due to the used disease model or the short duration of action with a single intravitreal injection of rhCNTF. On the other hand, the lack of in vivo efficacy was shown to not be due to distribution limitations; permeation into the retina was observed in both retinal explant models as in 24 h rhCNTF penetrated the inner limiting membrane, and being mostly observed in the ganglion cell layer, distributed to different layers of the neural retina. As rhCNTF can reach deeper retinal layers, in general, having direct effects on resident CNTF-responsive target cells is plausible.
Subject: retinal penetration
neuroprotection
protein aggregation
stability
intravitreal delivery
CNTF
CILIARY NEUROTROPHIC FACTOR
INTRAOCULAR GENE-TRANSFER
GANGLION-CELL SURVIVAL
INTRAVITREAL INJECTION
MOUSE-MODEL
PHOTORECEPTOR SURVIVAL
INTERLEUKIN-6 RECEPTOR
PROTEIN STABILITY
DRUG STABILITY
OUTER SEGMENTS
317 Pharmacy
Rights:


Files in this item

Total number of downloads: Loading...

Files Size Format View
pharmaceutics_12_00611_v2.pdf 7.001Mb PDF View/Open

This item appears in the following Collection(s)

Show full item record