Ádám Juhász

Bovine serum albumin-sodium alkyl sulfates bioconjugates as drug delivery systems.

Precipitation of bovine serum albumin (BSA) by anionic surfactants with alkyl chains of increasing lengths (octyl, decyl, dodecyl sulfates) was studied at room temperature, at pH 3.0, in isotonic sodium chloride solution. The particle size of albumin, the zeta potential, the surface charge and fluorescent properties of BSA-surfactant composites were investigated concerning addition of increasing amount of surfactant. The thermal stability of the systems was monitored by calorimetric analysis (DSC). The formation of the well-ordered structure in the self-assembly process in liquid phase was studied by XRD measurement. The structure of the precipitated BSA-surfactant nanocomposites was characterized by small-angle X-ray scattering (SAXS). Finally, ibuprofen (IBU) molecules were enclosed in BSA-surfactant bioconjugate systems and the release properties of the drug were investigated. It has been found out that, as a consequence to the increasing number of carbon atoms in the alkyl chains of the surfactant, the structure and the fluorescent properties of the aggregates formed can be controlled due to the increase in the hydrophobicity of BSA-surfactant composites. The bioconjugates are well applicable as carrier to realize controlled release of drug molecules.

Determination of binding capacity and adsorption enthalpy between Human Glutamate Receptor (GluR1) peptide fragments and kynurenic acid by surface plasmon resonance experiments.

The interaction between kynurenic acid (KYNA) and two peptide fragments (ca. 30 residues) of Human Glutamate Receptor 201-300 (GluR1) using surface plasmon resonance (SPR) spectroscopy was investigated. Because of the medical interest in the neuroscience, GluR1 is one of the important subunits of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPAR). AMPARs are ionotoropic glutamate receptors, which are mediating fast synaptic transmission and are crucial for plasticity in the brain. On the other hand, KYNA has been suggested to have neuroprotective activity and it has been considered for apply in therapy in certain neurobiological disorders. In this article the adsorption of the GluR1201-230 and GluR1231-259 peptides were studied on gold biosensor chip. The peptides were chemically bonded onto the gold surface via thiol group of L-cysteine resulted in the formation of peptide monolayer on the SPR chip surface. Because the GluR1231-259 peptide does not contain L-cysteine the Val256 was replaced by Cys256. The cross sectional area and the surface orientation of the studied peptides were determined by SPR and theoretical calculations (LOMETS) as well. The binding capability of KYNA on the peptide monolayer was studied in the concentration range of 0.1-5.0 mM using 150 mM NaCl ionic strength at pH 7.4 (±0.02) in phosphate buffer solutions. In order to determine the binding enthalpy the experiments were carried out between +10°C and +40°C. The heat of adsorption was calculated by using adsorption isotherms at different surface loading of KYNA on the SPR chip.

Kinetic and Thermodynamic Evaluation of Kynurenic Acid Binding to GluR1270-300 Polypeptide by Surface Plasmon Resonance Experiments

This work clearly demonstrates an evaluation process that is easily performed and is simply based on the fitting of temperature-dependent surface plasmon resonance (SPR) sensorgrams to provide detailed thermodynamic characterization of biologically relevant interactions. The reversible binding of kynurenic acid (KYNA) on human glutamate receptor (GluR1) polypeptide (GluR1270-300)-modified gold surface has been studied at various temperatures under physiological conditions by two-dimensional SPR experiments. The registered sensorgrams were fitted by using different kinetic models without application of any commercial software. Assuming that the association of GluR1270-300-KYNA complex is first order in both reactants, the association (ka) and dissociation (kd) constants as well as the equilibrium constants (KA) and the Gibbs free-energy change (ΔG°) were given at 10, 20, 30, and 40 °C. Moreover, the enthalpy (ΔH° = -27.91 kJ mol(-1)), entropy (ΔS° = -60.33 J mol(-1) K(-1)), and heat capacity changes (ΔCp = -1.28 kJ mol(-1) K(-1)) of the model receptor-ligand system were also calculated using a spreadsheet program. Negative values of ΔG° and ΔH° indicate the exothermic formation of a stable GluR1270-300-KYNA complex, because the |ΔH| > |TΔS| relation suggests an enthalpy-driven binding process. The negative ΔH° and ΔS° values strongly support the formation of a salt bridge between KYNA and the positively charged residues of the polypeptide (Arg, Lys) at pH 7.4, confirmed by molecular docking calculations as well.

Determination of binding capacity and adsorption enthalpy between Human Glutamate Receptor (GluR1) peptide fragments and kynurenic acid by surface plasmon resonance experiments. Part 2: Interaction of GluR1270–300 with KYNA

In the course of our previous work, the interactions of two peptide fragments (GluR1201-230 and GluR1231-259) of human glutamate receptor (GluR1201-300) polypeptide with kynurenic acid (KYNA) were investigated by surface plasmon resonance (SPR) spectroscopy. Besides quantitation of the interactions, the enthalpies of binding of KYNA on certain peptide fragment-modified gold surfaces were also reported. In the present work, a third peptide fragment (GluR1270-300) of the glutamate receptor was synthesized and its interaction with KYNA was investigated by an SPR technique. This 31-membered peptide was chemically bonded onto a gold-coated SPR chip via a cysteine residue. The peptide-functionalized biosensor chip was analyzed by atomic force microscopy (AFM) and theoretical calculations were performed on the structure and dimensions of the peptide on the gold surface. In order to determine the isosteric heat of adsorption of the binding of KYNA on the peptide-functionalized gold thin film, SPR experiments were carried out between +10°C and +40°C. The results on the GluR1270-300-KYNA system were compared with the previously published binding parameters of the interactions of GluR1201-230 and GluR1231-259 with KYNA. The binding abilities of KYNA with all three peptide fragments immobilized on the gold surface were estimated by a molecular docking procedure and the binding free energies of these AMPA receptor subunits with KYNA were determined.

Nucleotide-directed syntheses of gold nanohybrid systems with structure-dependent optical features: Selective fluorescence sensing of Fe3+ ions

This study demonstrates a one-step synthesis for the preparation of both adenosine monophosphate (AMP)-stabilized colloidal gold nanoparticles (AMP-Au NPs) and fluorescent gold nanoclusters (AMP-Au NCs). The dominant role of AMP:AuCl4- molar ratios in the formation of diverse nanosized Au products was proved. The size, the structure and the unique structure-dependent optical properties of the NPs and NCs were determined based on the results of numerous spectroscopic (UV-vis, fluorescence, infrared, x-ray photoelectron), high resolution electron microscopy (HRTEM) and dynamic light scattering (DLS) techniques. Stabile AMP-Au NPs with diameter of ca. 11nm and ultra-small AMP-Au NCs having blue fluorescence (λem=480nm) were identified. In addition, the AMP-Au NCs have been utilized to develop a selective sensor for the detection of Fe3+ ions in aqueous medium based on fluorescence quenching. Several essential metal ions and anions have been tested but our results clearly supported that dominant quenching was observed only for Fe3+ ions. Based on the determined limit of detection (LOD=2.0μM) our system is capable of detecting Fe3+ ions in drinking water. The Stern-Volmer constants (KSV) and various thermodynamic parameters (ΔG, ΔH°, ΔS°, ΔCp) of the quenching process have also been determined by the Stern-Volmer fitting of the fluorescence data in order to better understand the quenching mechanism.

Preparation of novel tissue acidosis-responsive chitosan drug nanoparticles: Characterization and in vitro release properties of Ca2+ channel blocker nimodipine drug molecules

ABSTRACT The pH‐responsive intelligent drug release facility of hydrophobically modified chitosan nanoparticles (Chit NPs) (d = 5.2 ± 1.1 nm) was presented in the case of poorly water soluble Ca2+ channel blocker nimodipine (NIMO) drug molecules. The adequate pH‐sensitivity, i.e. the suitable drug carrier properties of the initial hydrophilic Chit were achieved by reductive amination of Chit with hexanal (C6‐) and dodecanal (C12‐) aldehydes. The successful modifications of the macromolecule were evidenced via FTIR measurements: the band appearing at 1412 cm−1 (C–N stretching in aliphatic amines) in the cases of the hydrophobically modified Chit samples shows that the C–N bond successfully formed between the Chit and the aldehydes. Hydrophobization of the polymer unambiguously led to lower water contents with lower intermolecular interactions in the prepared hydrogel matrix: the initial hydrophilic Chit has the highest water content (78.6 wt%) and the increasing hydrophobicity of the polymer resulted in decreasing water content (C6‐chit.: 74.2 wt% and C12‐chit.: 47.1 wt%). Furthermore, it was established that the length of the side chain of the aldehyde influences the pH‐dependent solubility properties of the Chit. Transparent homogenous polymer solution was obtained at lower pH, while at higher pH the formation of polymer (nano)particles was determined and the corresponding cut‐off pH values showed decreasing tendency with increasing hydrophobic feature (pH = 7.47, 6.73 and 2.49 for initial Chit, C6‐chit and C12‐chit, respectively). Next the poorly water soluble NIMO drug was encapsulated with the C6‐chit with adequate pH‐sensitive properties. The polymer‐stabilized NIMO particles with 10 wt% NIMO content resulted in stable dispersion in aqueous phase, the formation of polymer shell increased in the water solubility/dispersibility of the initial hydrophobic drug. According to the drug release experiments, we clearly confirmed that the encapsulated low crystallinity NIMO drug remained closed in the polymer NPs at normal tissue pH (pH = 7.4, PBS buffer, physiological condition) but at pH < 6.5 which is typical for seriously ischemic brain tissue, 93.6% of the available 0.14 mg/ml NIMO was released into the buffer solution under 8 h release time. According to this in vitro study, the presented pH‐sensitive drug carrier system could be useful to selectively target ischemic brain regions characterized by acidosis, to achieve neuroprotection at tissue zones at risk of injury, without any undesirable side effects caused by systemic drug administration. Graphical abstract Figure. No Caption available.

Cross-linked and hydrophobized hyaluronic acid-based controlled drug release systems

This work demonstrates the preparation, structural characterization, and the kinetics of the drug release of hyaluronic acid (HyA)-based colloidal drug delivery systems which contain hydrophobic ketoprofen (KP) as model molecule. Because of the highly hydrophilic character of HyA the cross-linked derivatives at different cross-linking ratio have been synthesized. The hydrophobized variants of HyA have also been produced by modifying the polymer chains with cetyltrimethylammonium bromide (CTAB) at various HyA/CTAB ratios. Due to modifications the coherent structure of HyA changes into an incoherent colloidal system that were verified by rheological investigations. Nearly 70% of the encapsulated KP dissolve from the totally cross-linked HyA carrier but the release rate of KP is about 20% (after 8 h) from the CTAB-modified colloidal system at HyA monomer/CTAB 1:0.8 mass ratio. It has been verified that the modified HyA may be a potential candidate for controlled drug release of hydrophobic KP molecules.