Protamine-mediated efficient transcellular and transmucosal delivery of proteins

Proteins and peptides often require frequent needle-based administrations. Here, we report a non-parenteral delivery method for proteins through physical mixing with protamine, an FDA-approved peptide. Protamine was shown to promote tubulation and rearrangement of cellular actin, leading to enhanced intracellular delivery of proteins compared to poly(arginine)8 (R8). While the R8-mediated delivery resulted in significant lysosomal accumulation of the cargo, protamine directed the proteins to the nuclei with little lysosomal uptake. Intranasal delivery of insulin mixed with protamine effectively reduced blood glucose levels in diabetic mice 0.5 h after administration and the effect lasted for ∼6 h, comparable to subcutaneously injected insulin at the same dose. In mice, protamine was shown to overcome mucosal and epithelial barriers and modulate adherens junctions, promoting insulin penetration to the lamina propria layer for systemic absorption. [Display omitted] •Protamine increased intracellular delivery of proteins compared to R8 in 2D and 3D cell models.•Proteins delivered by protamine displayed cytosolic delivery and little accumulation in lysosomes.•Protamine enhanced intracellular delivery of proteins by promoting actin rearrangement of tubulation.•Protamine increased systemic absorption of proteins via intranasal delivery compared to R8.