Light-activated, in situ forming gel for sustained suprachoroidal delivery of bevacizumab.
Light-activated, in situ forming gel for sustained suprachoroidal delivery of bevacizumab.
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DOI：
10.1021/mp300716t
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发表时间：
2013-08-05
影响因子：
4.9
通讯作者：
Kompella UB
中科院分区：
文献类型：
--
作者：
Tyagi P;Barros M;Stansbury JW;Kompella UB
开发基于光活化聚己内酯二甲基丙烯酸酯和甲基丙烯酸羟乙酯的凝胶网络，该凝胶网络可持续释放稳定的活性贝伐单抗（一种用于治疗脉络膜新生血管的抗 VEGF 抗体），并评估在脉络膜上腔中原位凝胶形成后在兔眼中的持续离体递送和在大鼠眼中的体内递送。聚己内酯二甲基丙烯酸酯（PCM）由聚己内酯二醇（PCD）合成，并使用核磁共振波谱进行评估。 PCM 用于在 365 nm 紫外光和 2, 2-二甲氧基-2-苯基苯乙酮 (DMPA) 作为光引发剂的情况下交联甲基丙烯酸羟乙酯 (HEMA)。使用 3 分钟、7 分钟和 10 分钟 3 种不同的交联持续时间将贝伐珠单抗包埋在凝胶中。在为期 4 个月的研究中，使用基于 VEGF 结合的 ELISA 对贝伐珠单抗在 37°C PBS pH 7.4 中的体外释放进行了定量。通过尺寸排阻色谱（SEC）和圆二色性监测释放的贝伐单抗的稳定性。将与聚合物混合的 Alexa Fluor® 488 染料偶联贝伐单抗注射到兔眼脉络膜上腔，以研究不同交联持续时间对染料偶联贝伐单抗扩散的影响。通过注射与聚合物混合的 Alexa Fluor® 488 染料缀合贝伐单抗，然后交联 10 分钟，在 Sprague Dawley (SD) 大鼠中评估体内递送。使用眼底相机和 Fluorotron Master™ 无创监测在兔眼中的传播和在大鼠眼中的体内递送。通过NMR谱中羟基峰的消失证实了PCM的形成。 10 分钟的交联持续时间导致 21% 的贝伐单抗突释。其他交联持续时间的突释率≥ 62%。贝伐珠单抗从 10 分钟交联凝胶中的释放可持续约 4 个月。从 SEC 色谱图中观察到，释放样品中含有 ≥ 96.1% 的单体贝伐单抗。圆二色性证实贝伐珠单抗从凝胶释放后仍保持二级β-片层结构。随着交联持续时间增加到 10 分钟，凝胶/抗体更好地限制在切除的兔眼脉络膜上腔的注射部位。 Alexa Fluor® 488 染料结合贝伐单抗的递送在 SD 大鼠的脉络膜上腔中持续至少 60 天。 PCM和HEMA凝胶使贝伐单抗持续释放4个月，并保持了贝伐单抗的稳定性和VEGF结合活性。光激活 PCM 和 HEMA 凝胶适用于脉络膜上腔的原位凝胶形成和持续蛋白质递送。
To develop a light activated polycaprolactone dimethacrylate and hydroxyethyl methacrylate based gel network that sustains the release of stable, active bevacizumab (an anti-VEGF antibody used to treat choroidal neovascularization) and to assess sustained ex vivo delivery in rabbit eyes and in vivo delivery in rat eyes following in situ gel formation in the suprachoroidal space. Polycaprolactone dimethacrylate (PCM) was synthesized from polycaprolactone diol (PCD) and evaluated using NMR spectroscopy. PCM was used to cross-link hydroxyethyl methacrylate (HEMA) in the presence of 365 nm UV light and 2, 2-dimethoxy-2-phenylacetophenone (DMPA) as a photoinitiator. Bevacizumab was entrapped in the gel using 3 different cross-linking durations of 3, 7, and 10 minutes. In vitro release of bevacizumab in PBS pH 7.4 at 37°C during a 4 months study was quantified using a VEGF-binding based ELISA. Stability of released bevacizumab was monitored by size exclusion chromatography (SEC) and circular dichroism. Alexa Fluor® 488 dye conjugated bevacizumab mixed with polymers was injected suprachoroidally in rabbit eyes to study the effect of different cross-linking durations on the spread of the dye conjugated bevacizumab. In vivo delivery was assessed in Sprague Dawley (SD) rats by injecting Alexa Fluor® 488 dye conjugated bevacizumab mixed with polymers followed by cross-linking for 10 minutes. Spread in the rabbit eyes and in vivo delivery in rat eyes was monitored noninvasively using a fundus camera and Fluorotron Master™. Formation of PCM was confirmed by the disappearance of hydroxyl peak in NMR spectra. Cross-linking duration of 10 minutes resulted in a burst release of 21 % of bevacizumab. Other cross-linking durations had ≥ 62 % burst release. Bevacizumab release from 10 minute cross-linked gel was sustained for ∼ 4 months. Release samples contained ≥ 96.1 % of bevacizumab in the monomeric form as observed in SEC chromatograms. Circular dichroism confirmed that secondary β-sheet structure of bevacizumab was maintained after release from the gel. As the cross-linking duration was increased to 10 minutes, the gel/antibody was better confined at the injection site in excised rabbit eye suprachoroidal space. Delivery of Alexa Fluor® 488 dye conjugated bevacizumab was sustained for at least 60 days in the suprachoroidal space of SD rats. PCM and HEMA gel sustained bevacizumab release for 4 months and maintained the stability and VEGF-binding activity of bevacizumab. Light activated PCM and HEMA gel is suitable for in situ gel formation and sustained protein delivery in the suprachoroidal space.
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影响因子：
10.8
作者：
Koutsopoulos, Sotirios;Zhang, Shuguang
通讯作者：
Zhang, Shuguang
影响因子：
3.7
作者：
Cleland, JL;Jones, AJS
通讯作者：
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DOI：
10.1097/iae.0b013e3181d50cea
发表时间：
2010-06-01
影响因子：
3.3
作者：
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通讯作者：
Ammar, David A.
影响因子：
14
作者：
Cai, SS;Liu, YC;Prestwich, GD
通讯作者：
Prestwich, GD
影响因子：
3.9
作者：
Aishwarya, S.;Mahalakshmi, S.;Sehgal, Praveen Kumar
通讯作者：
Sehgal, Praveen Kumar