Making Oligonucleotides Better Biopharmaceuticals by Steric Protection

项目摘要/摘要 寡核苷酸面临几个生物制药方面的困难，包括稳定性和递送问题 作为非杂交活动，如凝血障碍和免疫系统的不受欢迎的激活。我们 已经开发出一种独特的寡核苷酸递送系统，称为pacDNA，它使用高密度 瓶刷聚合物提供具有结合选择性的寡核苷酸。这种聚合物相当于一个 熵障碍，减少了各种蛋白质对寡核苷酸的获取(从而产生副作用)，但仍然 允许不受阻碍的杂交。这种新的策略不仅提高了核酸酶的稳定性，保存了 靶结合能力，并最大限度地减少靶外副作用，但也大大增强了血浆 体内药代动力学、组织滞留和反义效力。我们目前的研究还显示， PacDNA的药理特性与瓶刷的脊椎密切相关。此外， PacDNA似乎是唯一能够逃避抗携带者适应性免疫的物质，这对 需要长期/频繁服药的疗法。最后，PacDNA沉积到组织和器官中。 缺乏成熟的输送技术，如皮肤、骨骼肌和心脏。这些 令人惊讶和令人信服的发现将成为下一个资助期调查的基础， 我们将1)使用具有以下特性的组合聚合物文库来探索pacDNA结构的属性空间 特定的骨架组成和单体序列；2)探针在体内的性质 小鼠和非人灵长类动物模型以及它如何能够逃避适应性免疫；以及3)探索 PacDNA有潜力创造一流/一流的疗法，利用其独特的优势 相关的临床前疾病模型(早衰症)。我们预计，这些目标的实现将 对这类材料有重要的基本理解，并使我们更接近临床 PacDNA的评价。

Project Summary/Abstract Oligonucleotides face several biopharmaceutical difficulties, including stability and delivery issues as well as non-hybridization activities such as coagulopathy and unwanted activation of the immune system. We have developed a unique oligonucleotide delivery system, termed pacDNA, which uses a high-density bottlebrush polymer to provide oligonucleotides with binding selectivity. The polymer amounts to an entropic barrier, reducing access to the oligonucleotide by various proteins (and thus side effects) but still allows for unhindered hybridization. This novel strategy not only improves nuclease stability, preserves target-binding capability, and minimizes off-target side effects, but also massively enhances plasma pharmacokinetics, tissue retention, and antisense potency in vivo. Our current studies also reveal that the pacDNA’s pharmacological properties are intimately related to the bottlebrush backbone. In addition, the pacDNA appears to be uniquely capable of evading anti-carrier adaptive immunity, which is useful for therapies that requires long-term/frequent dosing. Finally, the pacDNA deposits into tissues and organs that lack mature delivery technologies for, such as the skin, the skeletal muscle, and the heart. These surprising and enabling discoveries will be the basis for investigations in the next funding period, in which we will 1) explore the property space of the pacDNA structure using a combinatorial polymer library with specific backbone compositions and monomer sequences; 2) probe in vivo properties of the pacDNA in mouse and non-human primate models and how it is able to evade adaptive immunity; and 3) explore the potential of pacDNA to create first/best-in-class therapies that take advantage of its unique strengths using a relevant preclinical disease model (progeria). We anticipate that accomplishment of these objectives will yield significant fundamental understanding of this class of materials and bring us much closer to clinical evaluation of pacDNA.