Tuning the PEG surface density of the PEG-PGA enveloped Octaarginine-peptide Nanocomplexes

One of the main limitations of protein drugs is their restricted capacity to cross biological barriers. We have previously reported nanostructured complexes of insulin and modified octaarginine (C12-r8), enveloped by a polyethyleneglycol-polyglutamic acid (PEG-PGA) protective shell, and showed their capacity to overcome different barriers associated to the oral modality of administration. The objective of this work was to produce the said nanocomplexes with structurally diverse PEG-PGA shells, i.e. with different chain lengths and PEG substitution degrees, and comparatively analyze their PEG surface density and subsequent impact on their interaction with mucus glycoproteins and Caco-2 cells. The new PEG-PGA enveloped C12-r8-insulin nanocomplexes (ENCPs) exhibited a narrow size distribution (average size of 210-239 nm), a neutral surface charge and a 100% insulin association efficiency (final insulin loading of 16.5-29.6% w/w). Proton nuclear magnetic resonance (¹H NMR) analysis indicated the possibility to modulate the PEG density on the ENCPs from 6.7 to 44.5 PEG chains per 100 nm². This increase in the ENCPs PEG surface density resulted in their reduced interaction with mucins in vitro, while their interaction with Caco-2 cells in vitro remained unaltered. Overall, these data indicate the capacity to tune the surface characteristics of the ENCPS in order to maximize the capacity of these nanocarriers to overcome barriers associated to mucosal surfaces.