In-vitro permeation of bevacizumab through human sclera: effect of iontophoresis application
In-vitro permeation of bevacizumab through human sclera: effect of iontophoresis application
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DOI：
10.1111/j.2042-7158.2010.01153.x
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发表时间：
2010-09-01
影响因子：
3.3
通讯作者：
Nicoli, Sara
中科院分区：
文献类型：
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作者：
Pescina, Silvia;Ferrari, Giulio;Nicoli, Sara
目的贝伐珠单抗（阿瓦斯汀）是一种重组人源化单克隆抗体，用于眼科（标签外），用于治疗糖尿病视网膜病变和年龄相关性黄斑变性（湿性形式）等疾病中的新生血管形成。贝伐珠单抗目前采用重复玻璃体内注射的方式给药，这可能会导致严重的并发症；因此，需要一种非侵入性的递送途径。评估贝伐珠单抗通过分离的人巩膜的被动渗透，并探索离子电渗技术作为增强其体外经巩膜转运的方法。方法使用异硫氰酸荧光素（FITC）对贝伐珠单抗进行荧光标记。使用人巩膜作为屏障，在 Franz 型扩散池中进行 2 h 的渗透实验。供体隔室含有 FITC-贝伐单抗 (2.5 mg/ml)，溶于磷酸盐缓冲盐水，pH 7.4。在离子电渗实验中，施加的电流强度为2.3mA（电流密度3.8mA/cm2）。使用荧光检测器分析渗透样品（激发和发射波长分别为 490 和 520 nm）。通过薄层色谱检查 FITC-贝伐单抗缀合物的稳定性。 主要发现这项工作的主要发现是阳极离子电渗疗法可以显着增强贝伐单抗通过分离的人巩膜的转运（增强因子 7.5），即使该药物基本上不带电。由于抗体相对恒定的特性，这些结果可能会扩展到同一家族的其他分子。结论初步结果表明阳极离子电渗疗法可能是通过巩膜非侵入性输送贝伐珠单抗的一种有前途的策略。眼睛中存在其他静态和动态障碍，需要在更复杂的体外和体内模型上进一步评估该技术。
ObjectivesBevacizumab (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.MethodsBevacizumab 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 findingsThe 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.ConclusionsPreliminary 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.