Making Oligonucleotides Better Biopharmaceuticals by Steric Protection

项目摘要/摘要 使用寡核苷酸作为治疗剂有很大的兴趣，这面临着几个 生物制药困难，包括稳定性和递送问题，以及序列和/或化学物质 结构特定的、非杂交活动，如凝血病和免疫刺激 系统。这些困难在一定程度上被寡核苷酸的化学修饰所克服 或通过使用递送系统(通常是多阳离子结构)来增强核酸酶 稳定和提高交付效率。然而，这些方法要么带来了新的挑战 (例如毒性和免疫原性)，或不能充分解决所有负面方面。因此，a 一种可以提高核酸酶稳定性、保持靶标结合能力、最大限度减少脱靶 效果，改善生物分布仍然是非常受追捧的。我们的初步研究已经 证明了DNA的“紧凑”可以通过将其插入到高密度刷状聚合物中来实现 环境，使DNA能够选择性地结合到互补的DNA链上，同时访问 受到各种蛋白质的限制。DNA与核酸酶、Toll样受体9、 而凝血酶通常是非杂交性副作用的第一步。因此，刷子聚合物- DNA结合物应该会绕过寡核苷酸的许多副作用，并有可能成为 适用于几乎所有形式的寡核苷酸，即反义DNA、siRNA、microRNA、适体、 核酶等，以改善其生物制药特性。这项研究的结果将是 新型生物相容和非免疫刺激的基于寡核苷酸的基因治疗药物，以及 对其分子参数如何在体内和体外影响其的基本认识 属性。

Project Summary/Abstract Significant interests exist for using oligonucleotides as therapeutic agents, which face several biopharmaceutical difficulties, including stability and delivery issues, and sequence- and/or chemical structure-specific, non-hybridization activities, such as coagulopathies and stimulation of the immune system. These difficulties have been in part overcome by chemical modification of the oligonucleotide backbone or by using delivery systems (oftentimes polycationic structures), which enhance nuclease stability and improve delivery efficiency. However, these approaches either give rise to new challenges (e.g. toxicity and immunogenicity), or cannot adequately address all of the negative aspects. Therefore, a system that can improve nuclease stability, preserve target-binding capability, minimize all off-target effects, and improve biodistribution is still very much sought after. Our preliminary studies have demonstrated that “compaction” of DNA can be achieved by inserting it into a high-density brush polymer environment, which enables the DNA to bind selectively to a complementary DNA strand, while access by various proteins is limited. The binding of DNA with proteins such as nucleases, toll-like receptor 9, and thrombin is generally the first step to non-hybridization side effects. Therefore, the brush polymer- DNA conjugates should bypass many of the side effects of oligonucleotides and has the potential to be applied to essentially all forms of oligonucleotides, i.e. antisense DNA, siRNA, microRNA, aptamers, ribozymes, etc., to improve their biopharmaceutical characteristics. The outcome of this study would be a new class of biocompatible and non-immunostimulatory oligonucleotide-based gene therapy agents, and a fundamental understanding of how its molecular parameters can impact its in vivo and in vitro properties.