Fully modified bispecific aptamer for effective combination therapy of neovascular ocular diseases

项目摘要 糖尿病性黄斑水肿（DME）是工作年龄人群失明的主要原因。反- VEGF药物兰尼单抗和阿柏西普目前是DME的标准治疗。但这些 治疗远非最佳：约40%的DME患者对治疗反应不足。 患者必须无限期地接受定期玻璃体内注射，造成重大的治疗负担。 此后，又有两种抗VEGF药物在3期试验中进行了测试;尽管它们适度增加了 药物持续时间（从1-2个月到3个月），两者都未能进一步改善视力增益。因此，依靠反 单独的VEGF试剂显然不足以提高功效。 组合疗法，其将抗VEGF剂与靶向其它靶向的疾病调节剂组合联合收割机， 重要的生物标志物，如Ang 2和VEGF-C/D，在进一步提高疗效方面显示出巨大的前景 和/或减少治疗负担。然而，它们通常需要使用多种治疗剂， 有时需要多次注射，这进一步增加了成本和治疗负担。为了解决这一 缺点，理想的下一代DME处理应满足以下标准：a）体积小， 以实现高摩尔剂量和更长的功效持续时间，并促进组织渗透; B）缺乏 免疫原性和免疫刺激作用，以确保长期安全性; c）最重要的是， 能够用一种分子抑制多种生物标志物，以提高疗效而不增加 治疗负担和费用。 适体是单链寡核苷酸，其以类似于单克隆抗体的方式结合分子靶标。 抗体（mAb）。虽然适体的发明时间比单克隆抗体要晚得多，但它们已经 显示出作为眼部治疗剂的巨大潜力：Macugen，FDA批准的第一种抗VEGF药物， 湿性AMD治疗，是一种适体;另外两种适体（Fovista和Zimura）已在人体试验中进行了测试 对于多种视网膜适应症，使适体成为视网膜疾病最受评价的方式之一， 良好的安全性和明确的监管途径。 该SBIR的目的是开发高度稳定、多特异性的适体， DME的联合治疗。为此，Aptitude团队积累了丰富的经验， 适体发现我们以前开发的粒子显示方法，显着提高了 适体性能。我们还开发了筛选用于双特异性抗体的最佳接头的方法。 适体。此外，我们已经做了进一步的改进，直接筛选完全修饰的适体， 使药效持续时间更长。我们在适体发现方面的专业知识得到了我们合作者的补充。 DME临床前研究和临床试验的专业知识。如果成功，这个项目有可能带来 更有效和负担得起的治疗DME患者。

PROJECT ABSTRACT Diabetic Macular Edema (DME) is the leading cause of blindness in the working age population. The anti- VEGF agents Ranibizumab and Aflibercept are currently the standard of care for DME. However, these treatments are far from optimal: ~40% of the DME patients have an inadequate response to the treatment. Mover, patients have to receive regular intravitreal injections indefinitely, creating significant treatment burden. Two more anti-VEGF agents have since been tested in phase 3 trials; although they moderately increased drug duration (from 1-2 months to 3 months), both failed to further improve vision gain. Thus, relying on anti- VEGF agents alone is clearly insufficient to improve efficacy. Combination therapies, which combine an anti-VEGF agent with disease modifying agents that target other important biomarkers such as Ang2 and VEGF-C/D, have shown great promise in further improving efficacy and/or reducing treatment burden. However, they often require the use of multiple therapeutic agents and sometimes multiple injections, which further increase the cost and treatment burden. In order to address this shortcoming, the ideal next-generation DME treatment should meet the following criteria: a) small in size, in order to achieve high molar dosage and longer duration of efficacy, and facilitate tissue penetration; b) devoid of immunogenicity and immune-stimulatory effects, in order to ensure long-term safety; c) most importantly, capable of inhibiting multiple biomarkers with one molecule, in order to improve efficacy without increasing treatment burden and cost. Aptamers are single stranded oligonucleotides that bind to molecular targets in a manner similar to monoclonal antibodies (mAbs). Although aptamers were invented much more recently than mAbs, they have already shown significant potential as ocular therapeutics: Macugen, the first anti-VEGF agent approved by FDA for wet AMD treatment, is an aptamer; two more aptamers (Fovista and Zimura) have been tested in human trials for multiple retinal indications, making aptamers one of the most evaluated modality for retinal diseases, with a favorable safety profile and a clear regulatory pathway. The purpose of this SBIR is to develop highly stable, multi-specific aptamers that may serve as the optimal combination treatment for DME. To that end, the Aptitude team has accumulated extensive experience in aptamer discovery. We have previously developed the Particle Display method that significantly improves the aptamer performance. We have also developed the method to screen for the optimal linker for a bispecific aptamer. Moreover, we have made further improvement to directly screen for fully modified aptamers that may enable longer duration of efficacy. Our expertise in aptamer discovery is complemented by our collaborators’ expertise in DME preclinical research and clinical trials. If successful, this project has the potential of bringing more efficacious and affordable treatment to DME patients.