Novel polymeric nanoparticles for pulmonary nucleic acid therapy - synthesis, toxicological analysis and biological / therapeutic assessment

对于许多肺部疾病，包括肺肿瘤/转移，需要新的治疗策略。基于RNA干扰（RNAi）的治疗方法是有利的，但需要有效地递送小干扰RNA （sirna）。吸入siRNA应用具有直接接触、即时可用、有限的全身传播和较高的肺部剂量等优点，但面临重大挑战。基于聚合物或聚合物/脂质组合的纳米颗粒已被探索用于siRNA的体内递送。来自申请人和其他团体的初步数据表明，聚（乙烯）亚胺（PEIs）和其他多胺，以及针对定义的生物降解性和改进的NP稳定性的特定修饰，可能允许新型可吸入siRNA配方。除了疗效，毒性评估将关键性地决定肺给药的最佳NPs。此外，可吸入制剂的开发，例如基于雾化器或干粉吸入器，对临床应用至关重要。该项目将开发聚合物NPs作为输送癌基因特异性sirna的平台，通过吸入直接应用。这包括生成一个非常明确的、新型修饰的小多胺体系库，特别是酪氨酸修饰衍生物、可生物降解衍生物及其组合。新的NPs将在物理、物理化学、生物学和毒理学特性方面进行广泛的表征，并在肺转移小鼠模型中进行siRNA传递、生物分布和治疗效果的体内测试。不同的雾化策略（NP从水溶液或干粉，冻干或喷雾干燥后嵌入到水凝胶）将被探索。除了siRNA，开发的系统将扩展到微环质粒DNA和CRISPR-Cas9（即sgRNA + Cas9质粒）。最佳NPs的定义和生成，其吸入应用的最佳配方及其生物学评估依赖于DE和PL之间的高度相互作用，双方都贡献了其核心竞争力。在迭代过程中，基于新型聚合物（DE）的NPs的生物功效（DE， PL）和生物相容性/毒性（PL）的结果影响了聚合物修饰（DE）的进一步改进和NP的生成（DE， PL）。

For many pulmonary diseases including lung tumors / metastases, novel treatment strategies are needed. Therapeutics based on RNA interference (RNAi) are advantageous but require efficient delivery of small interfering RNAs (siRNAs). Inhalative siRNA application offers direct contact, immediate availability, limited systemic spread and thus higher doses in the lung, but meets major challenges.Nanoparticles based on polymers or polymer/lipid combinations have been explored for siRNA delivery in vivo. Initial data from the applicants’ and other groups indicate that poly(ethylene) imines (PEIs) and other polyamines, and specific modifications for defined biodegradability and improved NP stability, may allow for novel inhalable siRNA formulations. Beyond efficacy, the assessment of toxicity will critically determine optimal NPs for lung delivery. Additionally, the development of inhalable formulations, e.g. based on nebulizers or dry powder inhalers, is crucial for clinical applications. This project will develop polymeric NPs as a platform for the delivery of oncogene-specific siRNAs, for direct application via inhalation. This includes generating a library of very defined, novel modified small polyamine systems, esp. tyrosine-modified derivatives, biodegradable derivatives and combinations thereof. Novel NPs will be extensively characterized regarding physical, physicochemical, biological and toxicological properties, and tested in vivo in lung metastasis mouse models regarding siRNA delivery, biodistribution and therapeutic efficacy. Different nebulization strategies (NPs from aqueous solution or dry powder, lyophilized or spray-dried upon NP embedding into hydrogels) will be explored. Beyond siRNA, the systems developed will be extended towards minicircle plasmid DNA and CRISPR-Cas9 (i.e., sgRNA + Cas9 plasmid).The definition and generation of optimal NPs, their optimal formulation for inhalative application and their biological assessment rely on the high degree of interaction between DE and PL, with both sides contributing their core competences. In an iterative process, results on biological efficacy (DE, PL) and biocompatibility/toxicity (PL) of NPs based on novel polymers (DE) influence further refinement of polymer modifications (DE) and NP generation (DE, PL).