A novel vase-shaped molecular basket 1^3-, consisting of a deep nonpolar pocket of three napthalimide molecules fused to a bicyclic base, with polar glycine residues at the top was synthesized. Biocompatibility of the synthesized molecular basket was examined in vitro. The possible in vitro DNA protective potential of the newly synthesized deep-cavity basket 1^3- at several concentrations (25, 50, 100, 200, and 400 μg/mL) was investigated against hydroxyl and peroxyl radicals-induced DNA damage. Basket at lower concentrations (25, 50, 100 μg/mL) almost completely protected DNA against hydroxyl and peroxyl radicals induced DNA damage. It can be concluded that the newly synthesized molecular basket in the range of concentrations tested showed a significant DNA-protective potential against oxidative modifications of DNA caused by the harmful effects of free radicals.

The nose-to-brain route is one of the most promising alternative to promote drug delivery to the brain in the treatment of neurological diseases. Nasally administered drugs can be directly transported through the olfactory and trigeminal nerves, but enzymatic activity and the mucociliary clearance limit this process. Encapsulation of drugs in lipid nanoparticles, such as nanostructured lipid carriers (NLC), protects molecules against enzymatic activity, while promotes direct nose-to-brain transport. In this work, a valproic acid-loaded NLC (VPA-NLC) formulation was optimised using the quality-by-design (QbD) approach. A mixture design and a central composite design were used to optimise the critical material attributes (CMAs) and the critical process parameters (CPPs), respectively. The in vitro drug release profile and VPA-NLC morphology were investigated. The biocompatibility was assessed in human neuronal and nasal epithelial cells. VPA-NLC showed a particle size of 75 ± 1.05 nm, a polydispersity index (PDI) of 0.179 ± 0.006, an encapsulation efficiency (EE) of 85.7 % and a zeta potential (ZP) of 27.4 ± 0.351 mV. Transmission electron microscopy (TEM) images presented spherical nanoparticles smaller than 100 nm. Drug release studies showed about 50% of drug release after 6 hours and 100% after 24h. The VPA-NLC revealed safety up to 75 µg/mL in both cell lines. The optimised VPA-NLC formulation met the criteria of small particle size and PDI, and high EE and absolute ZP, which are required to follow the direct nose-to-brain transport. Additional experiments are being carried out to predict the in vivo safety and effectiveness of this formulation.

Ruta graveolens is a promising plant of traditional Ukrainian folk medicine for study. It is distributed in the southwestern part, namely in the Ivano-Frankivsk, Lviv, Transcarpathian region, on the Crimean Peninsula. In Ukrainian folk medicine, the plant is used for urolithiasis, women’s diseases, gastritis, nervous diseases, against convulsions, as an appetite stimulant. Also, rue has an effect on smooth muscles, the nervous system, and the structure of blood vessels. Ruta herb is included in the French and Brazilian pharmacopoeias, as well as into the British herbal pharmacopoeia. The plant is not included in the State Pharmacopoeia of Ukraine, and therefore requires detailed phytochemical analysis. The aim of our study was to identify the dominant marker compounds, which can be used to further standardization of herbal raw materials. Ruta herb was collected in the Ivan Franko National University of Lviv Botanical Garden (Ukraine) at July 2022. The plant powder (0.1 g) was extracted in 10 mL of 50% methanol in an ultrasonic bath (20 min) and then filtered. HPLC analysis of polypenolics was carry outed on Waters preparative HPLC Purification System with a Symmetry Prep C₁₈ (300×19mm×7µm) column, using the mobile phase composition of a mixture of 0.1% (v/v) trifluoroacetic acid in pure water (A) and acetonitrile (B) solvents. As a result, were determined neochlorogenic acid (0,03±0,01 mg/g), chlorogenic acid (0,11±0,01 mg/g), and rutin (6,09±0,25 mg/g). The research is still ongoing. Thus, the established composition indicates the prospects for further study and standardization of rue herb.

The hyperpolarization and cyclic nucleotide activated ion (HCN) channels have garnered increasing attention due to their association with the pathogenesis of various diseases. Given the heterogeneity in their expression patterns among different HCN channel subtypes, a comprehensive understanding of their function remains elusive and requires further investigation. The ion channel properties such as voltage dependence, are subject to modulation by endogenous cyclic nucleotides binding to the cyclic nucleotide binding domain (CNBD). In this study, we systematically assess the influence of seven distinct amino acids within the CNBD on ligand binding via the method of autodisplay and flow cytometry. Native HCN4 C-Linker-CNBD and its corresponding mutants were separately presented as fusion proteins on the surface of E. coli cells. Following incubation with 8-Fluo-cAMP, the whole cell fluorescence was quantified via flow cytometry. Perturbations in ligand binding, attributable to specific mutation, led to diminished fluorescence intensity (mFI) relative to the unaltered C-Linker-CNBD. Notably, mutations E660R, R669E and R710E resulted in a near-complete loss of whole cell fluorescence, confirming their importance for ligand binding as described before. Additionally, two amino acid substitutions, V642S and L652S, which were not anticipated to directly interact with the ligand, exhibited a strong reduction in mFI. We showed here for the first time the impact of these residues on ligand binding. Only moderate effects on ligand binding were observed for the mutants T670A and C662A. The presented ligand binding assay offers a rapid means of identifying residues essential for ligand binding. This information can be useful in the targeted drug design.

Interleukin (IL)-33, the newest member of the IL-1 family, plays a pivotal role in inflammatory and autoimmune diseases through its protein-protein interaction (PPI) with the ST2 receptor. Targeting this interaction holds promise for disease management. Although the IL-33/ST2 complex crystal structure has been resolved for nearly a decade, no comprehensive investigations into the druggability of IL-33 have been conducted. Furthermore, while several IL-33 inhibitors have been reported, their binding mechanisms have predominantly relied on rudimentary molecular docking approaches. In this study, we sought to identify possible druggable sites on the IL-33 surface using mixed-solvent molecular dynamics (MixMD) simulations and propose the possible mechanism of action of a reported IL-33 inhibitor using extensive-MD simulations. MixMD is an advanced MD technique that not only captures the protein's flexibility but also considers its interactions with small chemical probes. Our findings revealed five potential druggable sites on the IL-33 surface, two of which overlaid well with the interface of the ST2 receptor. The three remaining sites were investigated for their allosteric potential via all-atom normal mode analysis in the presence of pseudoligands. The current results suggested that interaction with these binding sites could exert possible dynamical change compared to the apoprotein conformation and serve as starting points for IL-33 allosteric modulation. Additionally, the binding modes of an orthosteric IL-33 inhibitor were also extracted and analyzed using the results from a 5-microseconds simulation. Our study can pave the way for future studies aiming to modulate the PPI of IL-33/ST2 employing both orthosteric and allosteric approaches.

Photopolymerization has emerged as a powerful technique for designing hydrogel-based transdermal delivery systems. This innovative method harnesses light-induced chemical reactions to create crosslinked hydrogel networks, offering precise control over drug release and enhanced therapeutic outcomes. This abstract explores the multifaceted role of photopolymerization in the formulation of transdermal hydrogel systems, discussing its principles, advantages, and diverse applications.

One of the primary advantages of photopolymerization is its ability to achieve rapid and on-demand gelation. This facilitates the incorporation of a wide range of therapeutic agents, including hydrophobic drugs and biologics, into the hydrogel matrix. Additionally, the spatiotemporal control afforded by photopolymerization enables the creation of gradient drug release profiles, optimizing drug permeation through the skin.

This abstract described challenges associated with photopolymerization-based hydrogel systems. The potential of photopolymerization to revolutionize the transdermal drug delivery field, by providing precise dosing control and minimizing skin irritation, makes it an exciting area of research in pharmaceutical and biomaterial sciences.

This research was carried out within the SMART-MAT Functional Materials Science Club of the Faculty of Materials Engineering and Physics of Cracow University of Technology as part of the 3rd edition of the program "Student research clubs create innovation" through the project titled „Transdermal systems in targeted therapy of skin cancer” financed by the Ministry of Science and Higher Education (grant no: SKN 157/568410/2023)

Biopolymeric hydrogels have gained significant recognition as versatile transdermal delivery systems in recent years. These hydrophilic three-dimensional networks, composed of natural polymers like chitosan, alginate, and hyaluronic acid, offer an innovative approach to enhance the percutaneous absorption of therapeutic agents. This abstract delves into the multifaceted role of biopolymeric hydrogels in transdermal drug delivery, discussing their formulation strategies, properties, and diverse applications.

One of the primary advantages of biopolymeric hydrogels is their ability to maintain a moist environment on the skin's surface, promoting efficient drug permeation while minimizing skin irritation. Their tunable physicochemical properties allow for controlled drug release, ensuring prolonged therapeutic effects and reduced dosing frequency. Moreover, these hydrogels can be tailored to encapsulate a wide range of drugs, including hydrophobic and hydrophilic compounds, proteins, and peptides.

This abstract also addresses the challenges associated with biopolymeric hydrogel-based transdermal delivery, such as optimizing drug release kinetics and ensuring long-term stability. However, their potential to revolutionize transdermal drug administration, particularly for chronic conditions, makes biopolymeric hydrogels a compelling area of research in pharmaceutical and biomedical sciences.

This research was carried out within the SMART-MAT Functional Materials Science Club of the Faculty of Materials Engineering and Physics of Cracow University of Technology as part of the 3rd edition of the program "Student research clubs create innovation" through the project titled „Transdermal systems in targeted therapy of skin cancer” financed by the Ministry of Science and Higher Education (grant no: SKN 157/568410/2023)

It is well known that studies of the luminescence of bases, nucleosides, nucleotides,
and especially their acid forms at room temperature are not sufficiently detailed concerning their
luminescence. Therefore, the aim of this study was to analyse and compare their spectral
properties. We studied the absorption and luminescence of monoribonucleotides and their
components dissolved in water at room temperature. For these samples, we measured absorption
spectra using a Specord 210plus instrument and excitation and emission of fluorescence and
phosphorescence spectra using Horiba Fluoro Max+ instruments. We observed a change in the
ratio between the peaks of the absorption spectra of acidic and salt forms of nucleotides. The
presence of luminescence was observed in nucleotides in both their acid and salt forms, as well as
in their nucleosides and bases. In addition, slight changes in the luminescence spectra were
observed for acid and salt forms of nucleotides. The luminescence intensity decreases from
nucleotides to bases. Therefore, our observations confirm that nucleotides, nucleosides, and
nucleic acid bases exhibit luminescence at room temperature, which may be useful information for
further research in this direction. In addition, changes in the luminescence spectra between acidic
and salt forms of nucleotides can be important diagnostic features in biochemical and medical
studies, where the identification and separation of different forms of nucleotides is important.

Parallel artificial membrane permeability assay (PAMPA) is a screening tool for the estimation of drug permeability across various biological membrane. We evaluated passive gastrointestinal absorption of fourteen new thiourea derivatives of naproxen using PAMPA test. Diffusion through artificial membranes that were composed of a mixture of hexadecane and hexane (the first PAMPA model) and of the solution of egg lecithin in dodecane (the second PAMPA model) was tested for the set of seven compounds and parent drug naproxen. The starting solutions were prepared by dissolving tested compounds in a phosphate buffer (pH 5.5), with the addition of DMSO (1%) and TWEEN80 (0.2%). These solutions were transferred into the donor PAMPA plates (300 µl) in triplicates. The acceptor solution (phosphate buffer (pH 7.4) and the same percentage content of DMSO and TWEEN as in the starting solutions) was transferred into the acceptor PAMPA plate (300 µl). Concentrations of analyzed compounds in the starting solutions, as well as in the donor and acceptor solutions after incubation were determined using an HPLC method. The first PAMPA model was selected for the estimation of passive gastrointestinal absorption of the remaining seven compounds. Derivatives 13 and 14 showed the lowest permeability coefficients (logPe values were -4.53 and -4.4, respectively). On the other hand, the highest permeability was determined for derivative 7 (logPe = -3.94). The highest membrane retention was observed for compound 6 (34%) and compound 3 (24%). This could explain lower permeability of these compounds than expected based on their lipophilicity.

Abstract: Cancer belongs to a group of diseases involving abnormal cell growth with the potential to spread to other parts of the body, being currently the leading cause of death worldwide. Cancer therapy is still very challenging and most of the traditional treatments available are usually invasive and cause serious side effects to patients. In this context, photodynamic therapy (PDT) emerges as a good alternative for the treatment of cancer, since it offers great benefits, such as low invasiveness and highly selective. This therapy combines a photosensitizer (PS), light of a specific wavelength, and molecular oxygen (³O₂) to produce reactive oxygen species (ROS), mainly singlet oxygen (¹O₂). Recent studies have shown that Coelenterazine (Clz) and its analogues can be used in PDT exhibiting relevant toxicity to different cancer cells (e.g., breast, liver, prostate, and neuroblastoma) without appreciable harm toward healthy cells. Synthetically, these coumpounds can be obtained from pyrazine units. To develop new photosynthesizers for the application in the PDT, herein we present the synthesis of new pyrazine-based scaffolds to be used for the preparation of novel PS. The different products were characterized by spectroscopy techniques.