Abstract
Objectives Bevacizumab (Avastin) is a recombinant humanized monoclonal antibody used in ophthalmology (off-label) for the treatment of neovascularization in diseases such as diabetic retinopathy and age-related macular degeneration (wet form). Bevacizumab is currently administrated by repeated intravitreal injection, which can cause severe complications; a non-invasive delivery route is therefore desirable. The passive permeation of bevacizumab through isolated human sclera was evaluated and the iontophoretic technique was explored as a method to enhance its transscleral transport in vitro. Methods Bevacizumab was fluorescently labelled using fluorescein isothiocyanate (FITC). Permeation experiments were conducted for 2 h in Franz-type diffusion cells using human sclera as the barrier. The donor compartment contained FITC-bevacizumab (2.5 mg/ml) in phosphate-buffered saline at pH 7.4. In the iontophoretic experiments, a current intensity of 2.3 mA (current density 3.8 mA/cm2) was applied. The permeation samples were analysed with a fluorescence detector (excitation and emission wavelengths were 490 and 520 nm, respectively). The stability of FITC-bevacizumab conjugate was checked by thin layer chromatography. Key findings The main finding of this work is that anodal iontophoresis can significantly enhance bevacizumab transport through isolated human sclera (enhancement factor 7.5), even though the drug is essentially uncharged. Due to the relatively constant characteristics of antibodies, these results can probably be extended to other molecules of the same family. Conclusions Preliminary results indicate that anodal iontophoresis could be a promising strategy to non-invasively deliver bevacizumab through the sclera. The presence in the eye of other barriers, both static and dynamic, necessitates further evaluation of the technique on more complex ex-vivo and in-vivo models.
| Original language | English |
|---|---|
| Pages (from-to) | 1189-1194 |
| Number of pages | 6 |
| Journal | Journal of Pharmacy and Pharmacology |
| Volume | 62 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - Sep 2010 |
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Keywords
- bevacizumab
- fluorescein isothiocyanate
- iontophoresis
- transscleral permeation
ASJC Scopus subject areas
- Pharmacology
- Pharmaceutical Science
Cite this
In-vitro permeation of bevacizumab through human sclera : Effect of iontophoresis application. / Pescina, Silvia; Ferrari, Giulio; Govoni, Paolo; MacAluso, Claudio; Padula, Cristina; Santi, Patrizia; Nicoli, Sara.
In: Journal of Pharmacy and Pharmacology, Vol. 62, No. 9, 09.2010, p. 1189-1194.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - In-vitro permeation of bevacizumab through human sclera
T2 - Effect of iontophoresis application
AU - Pescina, Silvia
AU - Ferrari, Giulio
AU - Govoni, Paolo
AU - MacAluso, Claudio
AU - Padula, Cristina
AU - Santi, Patrizia
AU - Nicoli, Sara
PY - 2010/9
Y1 - 2010/9
N2 - Objectives Bevacizumab (Avastin) is a recombinant humanized monoclonal antibody used in ophthalmology (off-label) for the treatment of neovascularization in diseases such as diabetic retinopathy and age-related macular degeneration (wet form). Bevacizumab is currently administrated by repeated intravitreal injection, which can cause severe complications; a non-invasive delivery route is therefore desirable. The passive permeation of bevacizumab through isolated human sclera was evaluated and the iontophoretic technique was explored as a method to enhance its transscleral transport in vitro. Methods Bevacizumab was fluorescently labelled using fluorescein isothiocyanate (FITC). Permeation experiments were conducted for 2 h in Franz-type diffusion cells using human sclera as the barrier. The donor compartment contained FITC-bevacizumab (2.5 mg/ml) in phosphate-buffered saline at pH 7.4. In the iontophoretic experiments, a current intensity of 2.3 mA (current density 3.8 mA/cm2) was applied. The permeation samples were analysed with a fluorescence detector (excitation and emission wavelengths were 490 and 520 nm, respectively). The stability of FITC-bevacizumab conjugate was checked by thin layer chromatography. Key findings The main finding of this work is that anodal iontophoresis can significantly enhance bevacizumab transport through isolated human sclera (enhancement factor 7.5), even though the drug is essentially uncharged. Due to the relatively constant characteristics of antibodies, these results can probably be extended to other molecules of the same family. Conclusions Preliminary results indicate that anodal iontophoresis could be a promising strategy to non-invasively deliver bevacizumab through the sclera. The presence in the eye of other barriers, both static and dynamic, necessitates further evaluation of the technique on more complex ex-vivo and in-vivo models.
AB - Objectives Bevacizumab (Avastin) is a recombinant humanized monoclonal antibody used in ophthalmology (off-label) for the treatment of neovascularization in diseases such as diabetic retinopathy and age-related macular degeneration (wet form). Bevacizumab is currently administrated by repeated intravitreal injection, which can cause severe complications; a non-invasive delivery route is therefore desirable. The passive permeation of bevacizumab through isolated human sclera was evaluated and the iontophoretic technique was explored as a method to enhance its transscleral transport in vitro. Methods Bevacizumab was fluorescently labelled using fluorescein isothiocyanate (FITC). Permeation experiments were conducted for 2 h in Franz-type diffusion cells using human sclera as the barrier. The donor compartment contained FITC-bevacizumab (2.5 mg/ml) in phosphate-buffered saline at pH 7.4. In the iontophoretic experiments, a current intensity of 2.3 mA (current density 3.8 mA/cm2) was applied. The permeation samples were analysed with a fluorescence detector (excitation and emission wavelengths were 490 and 520 nm, respectively). The stability of FITC-bevacizumab conjugate was checked by thin layer chromatography. Key findings The main finding of this work is that anodal iontophoresis can significantly enhance bevacizumab transport through isolated human sclera (enhancement factor 7.5), even though the drug is essentially uncharged. Due to the relatively constant characteristics of antibodies, these results can probably be extended to other molecules of the same family. Conclusions Preliminary results indicate that anodal iontophoresis could be a promising strategy to non-invasively deliver bevacizumab through the sclera. The presence in the eye of other barriers, both static and dynamic, necessitates further evaluation of the technique on more complex ex-vivo and in-vivo models.
KW - bevacizumab
KW - fluorescein isothiocyanate
KW - iontophoresis
KW - transscleral permeation
UR - http://www.scopus.com/inward/record.url?scp=77955242157&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77955242157&partnerID=8YFLogxK
U2 - 10.1111/j.2042-7158.2010.01153.x
DO - 10.1111/j.2042-7158.2010.01153.x
M3 - Article
C2 - 20796199
AN - SCOPUS:77955242157
VL - 62
SP - 1189
EP - 1194
JO - Journal of Pharmacy and Pharmacology
JF - Journal of Pharmacy and Pharmacology
SN - 0022-3573
IS - 9
ER -