Surfactant protein B-derived peptides as endosomal escape enhancers for pulmonary delivery of siRNA

Respiratory diseases still cause significant mortality and morbidity worldwide, highlighting the need for new inhalable drugs. RNA therapeutics, which have the potential to modulate the expression of virtually any gene, could address this unmet medical need. Nevertheless, clinical translation requires the design of RNA formulations able to overcome the extra- and intracellular barriers in the lung. We previously discovered that the endogenous cationic amphiphilic surfactant protein B (SP-B) promotes cytosolic delivery of small interfering RNA (siRNA) in lung-related cell types via endosomal membrane fusion. However, to bypass drawbacks related to the use of animal-derived SP-B, there is a keen interest in developing synthetic SP-B analogues with comparable activity. Here, we show that native SP-B can successfully be replaced by smaller peptides, with the N-terminal heptapeptide and amphipathic helix being minimally required to promote siRNA-induced gene silencing. Peptidolipid-coated nanogels were designed and demonstrated equivalent siRNA delivery efficacy compared to state-of-the-art lipid nanoparticles (LNPs). Moreover, they exhibit enhanced resistance to vibrating mesh nebulization and reduced inflammatory activation of bronchial epithelial cells. Collectively, the discovery of SP-B peptides as RNA delivery enhancers holds promise for developing potent inhalable RNA formulations with favorable safety profiles, of value for the treatment of chronic inflammatory pathologies.