Sustained Intravitreal Delivery of Ranibizumab Mediated by ECM Binding

项目总结 抗体治疗已成为许多眼科疾病的重要治疗手段。 血管形成，包括湿性老年性黄斑变性(AMD)，增殖性 糖尿病视网膜病变和脉络膜新生血管。阿瓦斯丁(贝伐单抗)和Lucentis(雷尼比珠单抗)分别为 人源化抗体及其Fab片段，分别通过阻断血管内皮生长因子来阻止血管生成。他们 在治疗这些眼病方面表现出显著的临床疗效；然而，目前的治疗方法 方案需要每月玻璃体内注射，因为药物通过快速扩散从玻璃体内清除。 眼睛。频繁注射导致严重的患者负担，增加了感染和反复注射的风险 与注射有关的压力和疼痛。延长抗体在眼睛中的活性将大大 提高治疗效用和疗效，减少毒副作用。此应用程序的总体目标是 开发能够以高亲和力结合玻璃体细胞外基质的雷尼比珠单抗(RAN)衍生物， 特别是通过与II型胶原(Coll)和透明质酸相互作用的工程多肽结合物 (HA)。长期目标是探索它们的持续释放，用于治疗黄斑变性和其他 眼球后部的疾病。中心假设是与玻璃体相互作用的RAN衍生物 玻璃体腔内注射ECM后，ECM的持续时间将显著延长，治疗活性也将得到改善。 在强劲的初步数据的指导下，这一假说将通过追求两个具体目标来检验。的关注点 目的1是合成能够与II型胶原或透明质酸(HA)结合的多肽-RAN结合物 维持其抗血管内皮生长因子的活性。结合物将测试细胞外基质结合亲和力和对血管内皮生长因子的抑制作用。 活性采用体外分子和细胞水平的分析。AIM 1还包括开发自组装的计划 可与多肽-RAN偶联物共注射的多肽纳米纤维，用于高水平的载药量 尤其是对于玻璃体中ECM水平较低的患者。目标2侧重于测试持续交付的 RAN配方和最终在延缓新生血管形成方面的长期疗效 诱导兔脉络膜新生血管模型。AIM 2还包括长期生物兼容性 RAN制剂的研究。这项提议意义重大，因为本地化和持续释放 通过细胞外基质结合和类细胞外基质纳米纤维制成的抗体药物可以应用于其他病理组织 富含胶原蛋白和透明质酸的皮肤和关节状况。

PROJECT SUMMARY Antibody-based therapies have emerged as an important treatment for many eye diseases with pathological blood vessel formation, including the wet form of aged related macular degeneration (AMD), proliferative diabetic retinopathy, and choroidal neovascularization. Avastin (bevacizumab) and Lucentis (ranibizumab) are humanized antibody and its Fab fragments, respectively, that prevent angiogenesis by blocking VEGF. They have shown remarkable clinical response in treating these eye diseases; however current therapeutic regimens require monthly intravitreal injections because of fast diffusion-mediated clearance of drugs from the eye. Frequent injections result in severe patient burden in terms of increased risk of infection and repeated stress and pain associated with the injection. Prolonging the activity of antibody in the eye would greatly improve the therapeutic utility and efficacy, and decrease toxicity. The overall objective of this application is to develop ranibizumab (Ran) derivatives that can engage the extracellular matrix of the vitreous with high affinity, particularly through engineering peptide conjugates that can interact with type II collagen (Coll) and hyaluronan (HA). The long-term goal is to explore their sustained delivery for treating macular degeneration and other disease of the posterior eye. The central hypothesis is that the Ran derivatives which interact with the vitreous ECM will have significantly extended duration and improved therapeutic activity after intravitreal injection. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims. The focus of Aim 1 is to synthesize peptide-Ran conjugates that can bind to type II collagen or hyaluronan (HA) while maintaining their anti-VEGF activity. The conjugates will be tested for ECM binding affinity and VEGF inhibiting activity using in vitro molecular- and cell-level assays. Aim 1 also includes plans to develop self-assembled peptide nanofibers that can be co-injected with peptide-Ran conjugates for high level of drug loading particularly for patients with low level of ECM in the vitreous. Aim 2 focuses on testing the sustained delivery of the Ran formulation and ultimately the long-term efficacy in delaying the neovasculature formation using induced choroidal neovascularization (CNV) model in rabbit. Aim 2 also includes the long-term biocompatibility study of the Ran formulations. The proposal is significant because localization and sustained release of antibody drugs made possible by ECM binding and ECM-like nanofibers can be applied to other pathologic conditions in the skin and joints which are rich in collagen and HA.