EPR Characterization of Copper(II) Complexes of PAMAM-Py Dendrimers for Biocatalysis in the Absence and Presence of Reducing Agents and a Spin Trap

Polyamidoamine (PAMAM) dendrimers at different generations (from G2 to G6) were functionalized with pyridine (Py) groups at the external surface, and their complexation behavior with Cu­(II) at increasing molar ratios between the ions and the Py groups was analyzed in the absence and presence of reducing agents and a spin trap. These Cu­(II)–dendrimer complexes may be used as antitumor and antiamyloidogenesis drugs, similarly to other Cu­(II)–dendrimer complexes, and as biocatalysts. Indeed, they have revealed to selectively catalyze molecular oxygen reduction to generate reactive oxygen species (ROS). A computer-aided electron paramagnetic resonance (EPR) study of these complexes allowed us to identify different complexes by increasing the Cu­(II)/Py molar ratio for the different generations. Binuclear EPR-silent complexes were formed at the highest generations. The differently complexed Cu­(II) ions showed a different capability to be reduced, starting from the most exposed at the dendrimer surface bearing a stable Cu­(II)–Py₂ coordination. Cu­(II)–G5 showed peculiar structural properties which probably favored its activity as biocatalyst. The spin trap was able to capture hydroxyl radicals, which became clearly EPR visible after all Cu­(II) ions were reduced to Cu­(I). This method may be used as a platform to study interactions of Cu­(II) in nanosized macromolecules for biomedical purposes, mainly in biocatalysis involving redox reactions and formation of ROS

Effect of Hydrogenated Cardanol on the Structure of Model Membranes Studied by EPR and NMR

Hydrogenated cardanol (HC) is known to act as an antiobesity, promising antioxidant, and eco-friendly brominating agent. In this respect, it is important to find the way to transport and protect HC into the body; a micellar structure works as the simplest membrane model and may be considered a suitable biocarrier for HC. Therefore, it is useful to analyze the impact of HC in the micellar structure and properties. This study reports a computer aided electron paramagnetic resonance (EPR) and ¹H NMR investigation of structural variations of cetyltrimetylammonium bromide (CTAB) micelles upon insertion of HC at different concentrations and pH variations. Surfactant spin probes inserted in the micelles allowed us to get information on the structure and dynamics of the micelles and the interactions between HC and CTAB. The formation of highly packed HC-CTAB mixed micelles were favored by the occurrence of both hydrophobic (chain–chain) and hydrophilic (between the polar and charged lipid heads) interactions. These interactions were enhanced by neutralization of the acidic HC heads. Different HC localizations into the micelles and micellar structures were identified by changing HC/CTAB relative concentrations and pH. The increase in HC concentration generated mixed micelles characterized by an increased surfactant packing. These results suggested a rod-like shape of the mixed micelles. The increase in pH promoted the insertion of deprotonated HC into less packed micelles, favored by the electrostatic head–head interactions between CTAB and deprotonated-HC surfactants

Effect of Hydrogenated Cardanol on the Structure of Model Membranes Studied by EPR and NMR

Hydrogenated cardanol (HC) is known to act as an antiobesity, promising antioxidant, and eco-friendly brominating agent. In this respect, it is important to find the way to transport and protect HC into the body; a micellar structure works as the simplest membrane model and may be considered a suitable biocarrier for HC. Therefore, it is useful to analyze the impact of HC in the micellar structure and properties. This study reports a computer aided electron paramagnetic resonance (EPR) and ¹H NMR investigation of structural variations of cetyltrimetylammonium bromide (CTAB) micelles upon insertion of HC at different concentrations and pH variations. Surfactant spin probes inserted in the micelles allowed us to get information on the structure and dynamics of the micelles and the interactions between HC and CTAB. The formation of highly packed HC-CTAB mixed micelles were favored by the occurrence of both hydrophobic (chain–chain) and hydrophilic (between the polar and charged lipid heads) interactions. These interactions were enhanced by neutralization of the acidic HC heads. Different HC localizations into the micelles and micellar structures were identified by changing HC/CTAB relative concentrations and pH. The increase in HC concentration generated mixed micelles characterized by an increased surfactant packing. These results suggested a rod-like shape of the mixed micelles. The increase in pH promoted the insertion of deprotonated HC into less packed micelles, favored by the electrostatic head–head interactions between CTAB and deprotonated-HC surfactants

Self-Assembled Ligands Targeting TLR7: A Molecular Level Investigation

Toll-like receptors (TLRs) are pattern recognition transmembrane proteins that play an important role in innate immunity. In particular, TLR7 plays a role in detecting nucleic acids derived from viruses and bacteria. The huge number of pathologies in which TLR7 is involved has led to an increasing interest in developing new compounds targeting this protein. Several conjugation strategies were proposed for TLR7 agonists to increase the potency while maintaining a low toxicity. In this work, we focus the attention on two promising classes of TLR7 compounds derived from the same pharmacophore conjugated with phospholipid and polyethylene glycol (PEG). A multidisciplinary investigation has been carried out by molecular dynamics (MD), dynamic light scattering (DLS), electron paramagnetic resonance (EPR), and cytotoxicity assessment. DLS and MD indicated how only the phospholipid conjugation provides the compound abilities to self-assemble in an orderly fashion with a maximal pharmacophore exposition to the solvent. Further EPR and cytotoxicity experiments highlighted that phospholipid compounds organize in stable aggregates and well interact with TLR7, whereas PEG conjugation was characterized by poorly stable aggregates at the cells surface. The methodological framework proposed in this study may be used to investigate, at a molecular level, the interactions generally occurring between aggregated ligands, to be used as drugs, and protein receptors

EPR and Rheological Study of Hybrid Interfaces in Gold–Clay–Epoxy Nanocomposites

With the aim to obtain new materials with special properties to be used in various industrial and biomedical applications, ternary “gold–clay–epoxy” nanocomposites and their nanodispersions were prepared using clay decorated with gold nanoparticles (AuNPs), at different gold contents. Nanocomposites structure was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Rheology and electron paramagnetic resonance (EPR) techniques were used in order to evaluate the molecular dynamics in the nanodispersions, as well as dynamics at interfaces in the nanocomposites. The percolation threshold (i.e., the filler content related to the formation of long-range connectivity of particles in the dispersed media) of the gold nanoparticles was determined to be ϕ_p = 0.6 wt % at a fixed clay content of 3 wt %. The flow activation energy and the relaxation time spectrum illustrated the presence of interfacial interactions in the ternary nanodispersions around and above the percolation threshold of AuNPs; these interfacial interactions suppressed the global molecular dynamics. It was found that below ϕ_p the free epoxy polymer chains ratio dominated over the chains attracted on the gold surfaces; thus, the rheological behavior was not significantly changed by the presence of AuNPs. While, around and above ϕ_p, the amount of the bonded epoxy polymer chains on the gold surface was much higher than that of the free chains; thus, a substantial increase in the flow activation energy and shift in the spectra to higher relaxation times appeared. The EPR signals of the nanocomposites depended on the gold nanoparticle contents and the preparation procedure thus providing a fingerprint of the different nanostructures. The EPR results from spin probes indicated that the main effect of the gold nanoparticles above ϕ_p, was to form a more homogeneous, viscous and polar clay–epoxy mixture at the nanoparticle surface. The knowledge obtained from this study is applicable to understand the role of interfaces in ternary nanocomposites with different combinations of nanofiller

Copper(II) Complexes with 4‑Carbomethoxypyrrolidone Functionalized PAMAM-Dendrimers: An EPR Study

The internal flexibility and interacting ability of PAMAM-dendrimers having 4-carbomethoxypyrrolidone-groups as surface groups (termed Gn-Pyr), which may be useful for biomedical purposes, and ion traps were investigated by analyzing the EPR spectra of their copper­(II) complexes. Increasing amounts (with respect to the Pyr groups) of copper­(II) gave rise to different signals constituting the EPR spectra at room and low temperature corresponding to different coordinations of Cu²⁺ inside and outside the dendrimers. At low Cu²⁺ concentrations, CuN₄ coordination involving the DAB core is preferential for G3- and G5-Pyr, while G4-Pyr shows a CuN₃O coordination. CuN₂O₂ coordination into the external dendrimer layer was also contributing to G3- and G4-Pyr spectra. The structures of the proposed copper–dendrimer complexes were also shown. G4-Pyr displays unusual binding ability toward Cu­(II) ions. Mainly the remarkably low toxicity shown by G4-Pyr and its peculiar binding ability leads to a potential use in biomedical fields