Making Oligonucleotides Better Biopharmaceuticals by Steric Protection

项目总结/摘要 寡核苷酸面临着一些生物制药的困难，包括稳定性和交付问题，以及 如非杂交活性，如凝血病和免疫系统的不需要的激活。我们 已经开发了一种独特的寡核苷酸递送系统，称为pacDNA，它使用高密度的 瓶刷聚合物以提供具有结合选择性的寡核苷酸。该聚合物相当于 熵屏障，减少各种蛋白质对寡核苷酸的访问（从而减少副作用），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， 靶向结合能力，并最大限度地减少脱靶副作用，但也大大提高了血浆 药物代谢动力学、组织保留和体内反义效力。我们目前的研究还表明， pacDNA的药理学性质与瓶刷骨架密切相关。此外该 pacDNA似乎是唯一能够逃避抗载体适应性免疫，这是有用的， 需要长期/频繁给药的治疗。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 令人惊讶和有利的发现将成为下一个资助期研究的基础， 我们将1）使用组合聚合物文库探索pacDNA结构的属性空间， 特定的骨架组成和单体序列; 2）探测pacDNA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创造一流/最佳疗法的潜力，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，实现这些目标将 产生对这类材料的重要基本理解，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.