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

项目摘要 糖尿病黄斑水肿(DME)是劳动年龄人群致盲的主要原因。反- 血管内皮生长因子制剂雷尼比珠单抗和阿普利赛特是目前治疗DME的标准药物。然而，这些 治疗远远不是最理想的：约40%的DME患者对治疗反应不足。 在这种情况下，患者必须无限期地接受定期玻璃体内注射，这造成了巨大的治疗负担。 自那以后，又有两种抗血管内皮生长因子药物在第三阶段试验中进行了测试；尽管它们的数量略有增加 用药时间从1-2个月到3个月，均未进一步提高视力。因此，依靠反 单靠血管内皮生长因子制剂显然不足以提高疗效。 联合疗法，将抗血管内皮生长因子制剂与针对其他疾病的疾病改良剂相结合 重要的生物标志物，如Ang2和VEGF-C/D，在进一步改善疗效方面显示出巨大的前景 和/或减轻治疗负担。然而，它们通常需要使用多种治疗剂和 有时多次注射，进一步增加了成本和治疗负担。为了解决这个问题 缺点是，理想的下一代DME处理应满足以下标准：a)体积小， 以达到高摩尔剂量和更长的有效时间，并促进组织渗透；b)无效 免疫原性和免疫刺激作用，以确保长期安全；c)最重要的是， 能够用一个分子抑制多个生物标志物，从而在不增加的情况下提高疗效 治疗负担和费用。 适配子是单链寡核苷酸，以类似于单克隆的方式与分子靶标结合。 抗体(单抗)。尽管适配子的发明比单抗要晚得多，但它们已经 显示出巨大的眼睛治疗潜力：Macugen，第一个被FDA批准用于 湿性AMD治疗是一种适配子；另外两种适配子(Fovista和Zimura)已经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评估最多的方式之一，具有 良好的安全性和明确的监管途径。 这种SBIR的目的是开发高度稳定的、多特异性的适体，这些适体可能是最佳的 二甲基醚的联合治疗。为此，智能团队在以下方面积累了丰富的经验 适配子的发现。我们之前已经开发了粒子显示方法，该方法显著改进了 适配子性能。我们还开发了一种方法来筛选双专一性的最佳连接子 适体。此外，我们还进一步改进了直接筛选完全修饰的适体的方法，这些适体可能 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的合作者的补充 在二甲基醚临床前研究和临床试验方面的专业知识。如果成功，这个项目有可能带来 为DME患者提供更有效和负担得起的治疗。

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.