Controlled delivery of the anti-VEGF aptamer EYE001 with poly(lactic-co-glycolic) acid microspheres
Controlled delivery of the anti-VEGF aptamer EYE001 with poly(lactic-co-glycolic) acid microspheres
复制标题
用聚乳酸-乙醇酸微球控制抗 VEGF 适体 EYE001 的递送
DOI：
10.1167/iovs.01-1156
复制
发表时间：
2003-01-01
影响因子：
4.4
通讯作者：
Adamis, AP
中科院分区：
文献类型：
--
作者：
Carrasquillo, KG;Ricker, JA;Adamis, AP
目的.目的：开发一种长效抑制血管内皮生长因子（VEGF）及其介导的效应的控释给药系统。通过油包油溶剂蒸发工艺开发了含有抗VEGF RNA适体（EYE 001）固态制剂的聚（乳酸-共-乙醇酸）（PLGA）微球。进行体外实验以表征释放曲线。通过监测RNA适体抑制人脐静脉内皮细胞（HUVECs）中VEGF诱导的细胞增殖的能力来测定释放的药物的稳定性和生物活性。用在短期、中期和长期释放时间点后收集的适体等分试样进行细胞增殖实验。为了研究这种聚合物装置作为潜在的经巩膜递送装置的可行性，开发了体外装置来评估聚合物通过兔巩膜的水合和降解以及随后通过其的递送。PLGA微球能够以持续的方式递送EYE 001，在20天的时间内平均速率为2 μ g/d。在冻干之前用二糖海藻糖固态稳定适体并包封在PLGA中使得药物在释放后更稳定。细胞增殖实验表明，释放后的适体的生物活性得到保留，如与VEGF孵育后抑制内皮细胞增殖所示。如扫描电子显微镜（SEM）所示，将微球装入密封室中并放置在兔巩膜的“眼眶”部分上6天，微球变得水合并开始降解。结果，如通过荧光定量法测定的，适体从微球通过巩膜递送。将含有适体的微球装载到装置中并将其放置在巩膜的眼眶表面上进行了评估，并显示是可行的。包封在PLGA中的RNA适体EYE 001在20天的时间内递送，保留活性。该方法代表了用于药物的经巩膜递送和脉络膜和视网膜疾病的治疗的有前景的方法。
PURPOSE. To develop a controlled-drug delivery system for the long-term inhibition of vascular endothelial growth factor (VEGF) and its mediated responses.METHODS. Poly(lactic-co-glycolic)acid (PLGA) microspheres containing anti-VEGF RNA aptamer (EYE001) formulations in the solid-state were developed by an oil-in-oil solvent evaporation process. In vitro experiments were performed to characterize the release profiles. Stability and bioactivity of the released drug were assayed by monitoring the RNA aptamer's ability to inhibit VEGF-induced cell proliferation in human umbilical vein endothelial cells (HUVECs). Cell proliferation experiments were conducted with aptamer aliquots collected after short-, mid-, and long-term release time points. To investigate the feasibility of this polymer device as a potential transscleral delivery device, an in vitro apparatus was developed to assess polymer hydration and degradation through rabbit sclera and subsequent delivery through it.RESULTS. PLGA microspheres were able to deliver EYE001 in a sustained manner, with an average rate of 2 mug/d over a period of 20 days. Solid-state stabilization of the aptamer with disaccharide trehalose before lyophilization and encapsulation in PLGA rendered the drug more stable after release. Cell proliferation experiments demonstrated that the bioactivity of the aptamer was preserved after release, as indicated by inhibition of endothelial cell proliferation after incubation with VEGF. Microspheres packed into a sealed chamber and placed onto the "orbital" part of a rabbit sclera for a period of 6 days became hydrated and started to degrade, as shown by scanning electron microscopy (SEM). As a result, the aptamer was delivered from the microspheres through the sclera, as determined spectrophotometrically.CONCLUSIONS. The loading of aptamer-containing microspheres into a device and placing it on the orbital surface of the sclera was assessed and shown to be feasible. RNA aptamer EYE001 encapsulated in PLGA was delivered over a period of 20 days with retained activity. This method represents a promising approach for the transscleral delivery of drugs and the treatment of choroidal and retinal diseases.