Stimuli-Responsive Directional Vesicular Assembly with Tunable Surface Functionality and Impact on Enzyme Inhibition

Abstract

The article describes the self-assembly of a series of unsymmetrical bola-shaped π-amphiphiles (NDI-1, NDI-1a, NDI-2, NDI-3, and NDI-4) consisting of a hydrophobic naphthalene-diimide (NDI) chromophore attached to a nonionic hydrophilic wedge and an anionic headgroup in the two opposite arms of the central NDI. By design, only a single hydrazide group is linked either on the ionic or nonionic arm of the NDI. NDI-1 and NDI-1a are regioisomers differing only in the location of the hydrazide group, placed in the nonionic or ionic arm, respectively. NDI-2, NDI-3, and NDI-4 are similar to NDI-1 in the placement of the hydrazide group but differ in the nature of the ionic headgroups. Except for NDI-2, all of them exhibit spontaneous vesicle structures in water (pH 9.0) as established by electron microscopy, small-angle neutron scattering, dynamic light scattering, and spectroscopy studies. Supramolecularly assembled oligo-oxyethylene chains of the hydrophobic wedge exhibited a lower critical solution temperature (LCST) at ∼40 °C, similar to that of covalent polymers. Consequently, above the LCST, the bola-amphiphile was converted to a single headgroup surfactant, resulting in the collapse of the vesicular structure to nanoparticles. In all examples, the dominant H-bonding force among the hydrazide groups resulted in unidirectional orientation, leading to the formation of a nonsymmetric membrane with the H-bonded chain located at the inner wall. Therefore, the functional group displayed in these vesicles could be fully dictated by the location of the hydrazide group. Thus, for NDI-1, NDI-3, or NDI-4, the hydrazide group, located at the nonionic arm, directed the nonionic wedge to converge at the inner wall of the vesicle by displaying the anionic headgroups toward the outer surface. In contrast, NDI-1a formed a nonionic vesicle because in this case anionic headgroups were located at the inner wall of the membrane. Furthermore, among NDI-1, NDI-3, and NDI-4, the charge density of the anionic surface and accordingly the radius of curvature and particle size could be tuned precisely as a function of the extent of charge delocalization in the phenoxide or carboxylate headgroup. These distinct self-assembly modes resulted in very different abilities of these vesicles for electrostatic-interaction-driven biomolecular recognition, which was studied by testing their ability to bind with cationic protein chymotripsin and inhibit its enzymatic activity. The enzyme inhibition ability followed the order NDI-1 > NDI-3 > NDI-4 > NDI-2 ≈ NDI-1a, which could be rationalized by their distinct functional group display and surface charge density factors.