Search of novel multifunctional ligands targeting neurodegereneration processes

1.Research project objectives/Hypothesis

The general aim of the project is to obtain novel multi-target directed ligands, which through inhibition of the b-amyloid production and aggregation and improvement of the cholinergic neurotransmission could ameliorate cognitive processes. Biological targets of planning studies are enzymes: acetylcholinesterase, butyrylcholinesterase, b-amyloid peptide and enzyme b-secretase, which are closely related to neurodegenerative processes and play an essential role in the pathogenesis and development of age-related dementia diseases, especially Alzheimer’s disease. Due to the complex pathophysiology of AD and identification of many potential targets, multiple ligand strategy will be applied in search for active agents. Its goal is to create a single molecule which can modulate multiple targets simultaneously (polypharmacology), with the aim of enhancing efficacy or improving safety relative to drugs that address only a single target.

2.Research methodology

Our project represents interdisciplinary researches including: molecular modeling, chemical

synthesis, biological assays and pharmacological tests. The starting point for this project are results of our recent studies and structures selected by using of molecular modeling technique. Chemical part of this project comprises development of synthetic pathways and synthesis new compounds: derivatives of N-benzylpiperazine, derivatives of isoindolo-1,3-dione and tetrahydro-1,3-isoquinolinedione (compounds I generation) and their analogs (compounds II generation). Biological activity of obtained compounds will be evaluated in vitro and in vivo. Their inhibitory potency against AChE/BuChE will be evaluated by using spectrophotometric Ellman’s method. BACE1 in vitro activity and inhibition will be measured by using fluorescence resonance energy transfer (FRET) technology. Inhibition of Ab aggregation and formation will be tested by using a Thioflavin T fluorescence method. Investigations de novo in silico molecular modeling will be included in the second phase realization of the project – designing of analogs multifunctional ligands (compounds II generation). Subsequently, for selected leads (six compounds) pharmacological evaluation will be performer including learning and memory tests: novel object recognition, passive avoidance and Barnes maze test and general behavior studies: locomotor activity, rotarod and chimney test.

3.Research project impact

As the results of the project we are expecting to obtain novel, bioactive, multifunctional compounds with b-secretase inhibitory, cholinesterases inhibitory and b-amyloid anti-aggregating activities, therefore essential for disease-modifying role in the treatment of Alzheimer’s disease and also important for symptomatic treatment. Outcome of the project will be a library of new, original chemical compounds, derivatives of Nbenzylpiperazine, derivatives of isoindolo-1,3-dione and tetrahydro-1,3-isoquinolinedione and their analogs. Performed in vitro biological tests could give us the knowledge of biological properties of these molecules and pharmacological tests could estimate their precognitive properties. Investigations of novel molecules could help us to understand the mode of action of these compounds as well as could be a source of new knowledge on biology and pathophysiology of Alzheimer’s disease.