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No description available.

Global disease burden due to age related cognitive decline and dementias, especially Alzheimer’s disease, are a growing public health problem. Current treatments fail to completely prevent or cure dementia. A target of interest is the Insulin Regulated Aminopeptidase (IRAP) enzyme, inhibitors of IRAP have shown promise in preventing and reversing neurological degeneration associated with dementia. Structural information about IRAP when bound to an inhibitor is lacking, however, and future ligand design depends on knowledge of the mechanism of inhibition. We used long term MD simulations of IRAP bound to various ligands to investigate the conformational changes undergone by IRAP. PCA analysis was also used to investigate larger changes in the enzyme. A highly stable pose for a known spiro-oxindole inhibitor was found with key interactions between the ligand and SER546 and TYR 954, providing useful data for the design of future inhibitors of the IRAP enzyme.

Psilocybin is a hallucinogenic compound found in fungi and is currently evaluated inclinical trials for treatment of depression, anxiety and addiction. Psilocybin is aprodrug of the pharmacologically active metabolite, psilocin. The synthetic route topsilocybin relies on synthesizing psilocin from the starting material, 4-hydroxyindoleand latter converting psilocin into psilocybin by phosphorylation. The synthesis ofpsilocybin has been challenging because of the labile nature of the phosphorylationdibenzyl ester reagent and related intermediates. Several attempts to optimizepsilocybin synthesis have been published but there is still a need for furtherimprovements.The first aim of this project was to synthesize psilocybin using literature methods,while the second aim was to optimize the phosphorylation step with differentreagents and conditions.Initial studies focused on coupling the two-side chain carbon onto position three ofthe indole, this required protection of the hydroxyl group which was achieved byacylation in room temperature. With sufficient amount of dimethylamine solution, theamine addition reaction was investigated and resulted in a pure product. Thefollowing reduction with lithium aluminum hydride provided an unknown side-productinstead of psilocin.The first aim was successfully accomplished, up to psilocin, with pure intermediatesbut low yields. The second aim was not achieved due to lack of time and access tothe laboratory during covid-19 crisis. However, a literature survey of reagents andconditions for phosphorylation was performed which enables continuation of theproject.

No description available.

No description available.

Adenosine receptors (ARs) are a family belonging to the GPCR superfamily, involved with multiple physiological processes and widely distributed throughout the body. In this study, we focus on the A3 AR due to its vast scale applications in various pathophysiological conditions such as inflammation, pain, allergic asthma, and cancer chemotherapy. We report the validation of the binding mode of A3 AR antagonists, through a comprehensive in-silico mutagenesis study using free energy perturbations, reproducing prior experimental site-directed mutagenesis data for A3 AR antagonists. After validating the antagonist binding mode, we performed an extensive in-silico site-directed mutagenesis study on the hA3 AR using potent, selective, and structurally simple A3 AR antagonist based on the N-(2,6-diarylpyrimidin-4- yl)acetamide scaffold (pyrimidine core). The results of this study will be used to design in- vitro site-directed mutagenesis performed by collaborators (University of Barcelona). Once the binding mode of this scaffold is validated, it will be the basis for the design of compounds with two well-defined functionalities: a fluorophore moiety and bivalent ligands that target A3 dimers. The discovery of novel mutations on the hA3 AR is a step forward in the development of both chemically simple, potent, and selective A3 AR antagonists as well as in the characterization and crystallization of the A3 AR.

Subcutaneous administration of biological drugs has become highly attractive as it offers the possibility for patient self-administration. Coacervates as a subcutaneous drug delivery system provide a way to decrease injection volumes thereby reducing the risk of injection site pain. The aim with this thesis was to investigate possible coacervation between polyelectrolytes and peptide drugs under various physicochemical conditions. In this project, hyaluronic acid (HA) and carboxymethylcellulose (CMC) and their ability to coacervate with polymyxin B (PB) and vancomycin (VA) was studied. Furthermore, the release of these peptide drugs from the formulations into a release medium mimicking the subcutaneous environment was studied using UV spectroscopy. Studies showed successful coacervation between VA-CMC, PB-CMC and PB-HA. VA-CMC and PB-CMC coacervates were formed at higher peptide-polyelectrolyte charge ratios and lower ionic strengths at pH 7. The increase in charge ratio seemed to eliminate steric effects caused by the polyelectrolyte chains that most likely hindered coacervation. Furthermore, PB-CMC and PB-HA formed gel-like coacervates in the pH range of 11.73-11.84 at 1:1 charge ratio and ionic strengths 0-70mM. At such high pH values, the obtained formulations were most likely a result of PB-PB aggregation or charge regulation. From drug release studies one could conclude that VA-CMC exhibits direct release followed by a slower prolonged release profile. PB-CMC coacervates, liquid and gel, showed a sustained release profile while the PB-HA gel formulation resulted in direct release. Unfortunately, due to limitations with the experimental release rate set-up, the conclusions drawn can not be considered 100% reliable.

Fragment-based virtual screening is an in-silico method that potentially identifies new startingpoints for drug molecules and provides an inexpensive and fast exploration of the relevantchemical space compared to its experimental counterpart. It focuses on docking small potentialbinding fragments to a binding pocket and is used to design improved binders by growing thefragments or joining fragments using suitable linkers. In this project, a fragment-based virtualscreening was evaluated by docking 21 fragments that are obtained from 4 different drugs. Here,the fragments were evaluated using SP score in place and SP and XP flexible docking methodsand were compared to the results of the two decoy fragment datasets. Three of the investigatedfragments are positioned at the top and docked with the correct poses and pockets when comparedto the corresponding substructure in the crystal structure and thus could be considered a successfulfragment starting points. Out of the two flexible docking methods used, the SP method providedadditional correct poses and pockets than XP in this limited dataset.

Cancer remains a challenge in modern medicine, requiring the continuous search for new therapeutic interventions. Malic enzyme 2 (ME2), involved in cellular energy metabolism and redox balance, has appeared as a potential target for cancer therapy due to its overexpression in various cancer types. The aim of this project is to screen and validate compounds with ME2-inhibitory activity using a combination of computational modeling and enzymatic assays. A pharmacophore model was constructed and used to screen a database of compounds in order to generate potential ME2 inhibitors. The inhibitory effect on ME2 of five compounds was evaluated by measuring the formation of NADH using a spectrophotometer. Dose-response curves were plotted and the IC50 values were determined, revealing that these compounds have an inhibitory effect on ME2. Future studies should focus on potential sources of error and optimize conditions to ensure reproducibility and accuracy in assessing the inhibitors. / Cancer förblir en utmaning och kräver kontinuerlig forskning kring nya terapeutiska strategier. Malatdehydrogenas 2 (ME2) som är inblandad i cellens ämnesomsättning och redoxbalans har framträtt som en potentiell måltavla för cancerbehandling på grund av att den överuttrycks i olika cancerformer. Syftet med detta projekt är att testa och validera föreningar med ME2-hämmande aktivitet m.h.a en kombination av datorbaserad modellering och enzymatiska analyser. En farmakoformodell skapades utifrån kända ME2 inhibitorer och användes för att undersöka en databas med föreningar, i syfte att generera potentiella inhibitorer. Den hämmande effekten på ME2 i fem föreningar utvärderades genom att mäta bildningen av NADH med en spektrofotometer. “Dose- response” kurvor skapades och IC50 -värdena bestämdes. Resultaten visade att dessa föreningar har en hämmande effekt på ME2. Framtida studier bör fokusera på potentiella felkällor och att optimera de experimentella förhållandena för att säkerställa reproducerbarhet och ytterligare bekräfta föreningarna.