Preparation and in vitro testingof HLö7 - universal reactivator of inhibited acetylcholinesterases
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
David Malinak, Jan Korabecny, Miroslav Psotka, Martina Hrabinova, Jana Janockova
Abstract:
The aim of the project is in particular to develop and implement the simpler synthetic pathway required for the preparation of compound HLö7 and also in vitro testing of the prepared substance. As is currently known, for the synthesis of HLö7 is use a compound known as bis(methylsulfonoxymethyl)ether, the synthesis of which is difficult. This requires special instrumentation (eg very low pressure distillation, etc.). Furthermore, HLö7 can not be prepared by known multi-steps methods in amounts required for in vivo testing, and known procedures are often not reproducible. Based on these findings, the main aim of the project is to optimize the conditions and thus to simplify of the preparation of this most effective reactivator of AChE. The aim of the project will also in vitro testing of substance to determine the inhibition of AChE, and also the verify of reactivation activity on selected pesticides (dichlorvos and paraoxone) and to determine cytotoxicity on selected cell lines.
Project duration:
1-January-2017 - 31-December-2017
N-Propargyl-naphthoquinone hybrids: Development of novel potential drugs to combat Alzheimer’s disease
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Jan Korabecny, Eva Mezeiova, Katarina Spilovska, Petr Jost, Martina Hrabinova, Jana Janockova
Abstract:
The aim of the proposed project is to synthesize novel hybrid compounds capable to inhibit amyloid beta aggregation. Moreover, these compounds are predicted to decrease the burden caused by oxidative stress via MAO-B inhibition. Additionally, MAO-A inhibition will confer antidepressant and anxiolytic effects, symptoms accompanying AD. Such multi-target-directed ligands, chemically derived from naphthoquinone with attached N-propargylamine moiety will be further in vitro tested to validate their real therapeutic potential. Based on the obtained results, naphthoquinone-N-propargylamine hybrids will serve as new leads to AD treatment.
Project duration:
1-January-2017–31-December-2017
Analysis of non-bonding interactions between small molecules and cathepsin B by HDX mass spectrometry
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Rafael Dolezal, Jana Zdarova Karasova, Karel Musil, Martin Novak, Marketa Benkova
Abstract:
The aim of the project is to introduce a new and modern analytical methodology in our workplace, which is focused onto molecular research of potential pharmaceuticals. The HDX-MS method is a world-wide recognized alternative to X-ray and NMR, although its application, optimization and correct interpretation of the results obtained is of course not easy. Along with the development of precise technology for mass spectrometry (MSE, ion mobility, bioinformatics), HDX-MS represents a perspective method that allows to analyze the molecular protein structure, dynamics, conformational flexibility, protein-protein or protein-ligand interactions for samples of subnanomolar quantities. At present, the proteomic shot-gun analysis and the determination of metabolites of the first phase of in vitro metabolism by LC-MS / MS are very well developed at the CBV site, which is an important basis for successful handling of HDX-MS techniques. The present project therefore aims to participate in the comprehensive research and development of new drugs at CBV FNHK, especially against Alzheimer's disease and cancer. As a biological target, cathepsin B was selected for this project. Within the present project, complexes of 5 selected low molecular modulators with cathepsin B will be analyzed using the HDX-MS method.
Project duration:
1-January-2017–31-December-2017
In vitro prediction of the permeability of potential drugs using a MDCK cell line
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Ondrej Soukup, Rafael Dolezal, Jana Janockova, Marketa Benkova, Tereza Kobrlova
Abstract:
The primary aim of the project is to implement a new methodology, representing a "living" model of the blood-brain barrier represented by the MDCK cell line. The methodology will be validated by means of standard drugs in the treatment of AD (e.g., donepezil) and substances not passing the blood-brain barrier (e.g. atenolol). Active transport and efflux mechanisms of this line will be investigated by standard drugs of active transport or are substrates of efflux transporters such as P-gp (ABCB1) and so called multidrug resistance - associated protein 1 (ABCC1) and their inhibitors (LY335979, MK-571) .
Project duration:
1-January-2017–31-December-2017
Expression and purification of human recombinant beta secretase and validation in vitro assay for the determination its activity
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Martina Hrabinova, Monika Schmidt, Daniel Jun, Kamil Musilek, Vendula Sepsova
Abstract:
The main aim of this project is expression and purification of recombinant protein beta secretase (BACE1), which might be potential strategy for Alzheimer’s disease therapy.Recombinant human BACE1 will be producedvia the human embryonic kidney 293(HEK293) cell culture, which ensure all necessary post-translational modification of the protein. This setup produce enzyme with high activity and stability, which is need for validation of the colorimetric assay. This new method upgrade portfolio of enzymes assay participated in Alzheimer’s disease pathophysiology at our department. Based on the achieved results, GACR or AZV project is going to be submitted.
Project duration:
1-January-2017–31-December-2017
New compounds affecting pattern recognition receptors as potential adjuvants/immunomodulatoes
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Jan Honegr, David Malinak, Rafael Dolezal, Marketa Benkova
Abstract:
The aim of the project is to identify novel potential ligands of TLR4 with an internal activity comparable with standard - MPLA by in vitro high-throughput screening on cell lines stably co-expriming human TLR4. Novel compounds will be designed by the computer assisted molecular modeling with utilization of the molecular docking. The selected hit compounds will be synthesized and tested for cytotoxicity and internal activity. Simultaneously we will be screening compounds from BRC database to found if some of them possess internal activity on TLR.
Project duration:
1-January-2017–31-December-2017
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Juraj Lenco, Marie Vajrychova , Marketa Benkova
Abstract:
The project aims to further improve the high-flow LC-MS analyses exploited in our laboratory, so that their productivity could compete with the standard but unfortunately less robust nanoLC-MS analyses. We will deal with phenomena that affect both, the chromatographic efficiency and MS productivity such as peak dispersion, thermal mismatch of mobile phases entering the HPLC column, ionization efficiency, and mobile phase acidifiers. Only thoroughly optimized and thus sufficiently-yielding LC-MS proteomic analyses, have a chance to succeed in clinical settings where the emphasis is on speed, robustness and stability.
Project duration:
1-January-2017–31-December-2017
Establishing of new methods for in vitro testing of potential drugs Alzheimer´s diseases
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Vendula Sepsova, Daniel Jun, Martina Hrabinova, Ondrej Soukup, Jana Janockova
Abstract:
New two screening methods will be established. These methods enable evaluating new substances in vitro, which affecting major therapeutic targets of AD. The primary aim is to developed an alternative methodology for measuring aggregation Aβ, its inhibition respectively. Thus, a new spectrophotometric method will be implemented. Kongo Red will be used as the main agent, besides it is used in histopathology for amyloidosis diagnosis. It was proved, that Aβ aggregation depends on an ability of AChE inhibitor binding into the enzyme PAS. It is possible to test this interaction thanks to second established test (ThT). The secondary aim is to measure (after successful validation of new methods) group of selected AChE inhibitors.
Project duration:
1-January-2017–31-December-2017
Extension of in vitro tests for the evaluation of potential therapeutic of neurodegenerative drugs
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Ondrej Soukup, Rafael Dolezal, Jana Janockova, Marketa Pasdiorova, Eugenie Nepovimova, Jan Korabecny
Abstract:
The primary objective is to implement a new methodology for tjhe workplace. Historically our work focused on the development and testing of new drugs in accordance with cholinergic theory of treatment (inhibition of AChE interaction with muscarinic receptors). Recently, we have established a methodology to assess the inhibition of beta-secretase or prolyl oligopeptidases, which proved to be active for collaboration and the ensuring publication outputs. The newly proposed tests (inhibition of GSK-3beta, antiplatelet activity against Abeta 1-24 and the interaction with nicotinic receptors), thus broadening the existing range of tests and know-how of the workplace, so that it can be utilized in the search for and development of neovel drugs. These new tests will then be offered in the form of cooperation between research worplaces and private companies in the form of services. The secondary objective is to test the new range of potential drugs on mentioned antiplatelet activity and inhibit Abeta GSK3beta.
Project duration:
1-January-2016 - 31-December-2016
Cyclophilin D (CypD) as a potential target for therapeutic intervention within Alzheimer´s disease
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Kamil Musilek, Ondrej Benek, Lukas Hroch, Martina Hrabinova, Daniel Jun
Abstract:
The project is aimed to evaluation of cyclophilin D (CypD) as a potential target for therapeutic intervention within Alzheimer´s disease. For this purpose, the novel compounds affecting CypD activity will be prepared and evaluated in vitro with standards (e.g. cyclosporin A, antamanide). The novel method for CypD evaluation will broaden the pool of mitochondrial targets that are assumed to take part in progression of Alzheimer´s disease, and it will allow to gain preliminary data for further external project.
Project duration:
1-January-2016 - 31-December-2016
Development of novel telangiectasia and Rad3-related protein kinase (ATR) inhibitors derived from caffeine for their potential use in cancer treatment
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Jan Korabecny, Martin Andrs, Lukas Gorecki, Ales Tichy, Rafael Dolezal, Eugenie Nepovimova
Abstract:
The aim of the current project is to develop potential ATR inhibitor. Our knowledge in this field of drug discovery stems from the well-known caffeine ATR inhibition properties which is slightly selective to ATR kinase. Having these in mind, we will utilize common synthetic approaches to develop novel water-soluble derivatives in order to improve selective profile of novel caffeine derivatives to ATR over other kinases. These analogues will be subsequently tested in vitro in collaboration with Faculty of Medicine in Hradec Kralove, Charles University and Faculty of Military Health Sciences, University of Defense. As a result, structure-activity relationship will be established. The study also includes cytotoxicity assessment on different carcinoma cell lines. The most perspective hybrids will be further evaluated in ongoing preclinical studies.
Project duration:
1-January-2016 - 31-December-2016
Quaternary ammonium salts as a new disinfection means
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Ondřej Soukup, PhD, Lenka Ryšková, MD, Lenka Hobzová, MD., Daniel Jun, PhD., Dávid Maliňák, PhD, Šárka Salajková Ing.
Abstract:
The aim of this project is to prepare the 20 new, unpublished, substances from the group of water-soluble quaternary ammonium salts (QAS) with a high disenfection potential against Gram-negative and Gram-positive bacteria and against yeast and filamentous fungi. Then to select most effective candidates and mix them in order to prepare a product that would cover the entire spectrum of the tested pathogens (efficacy). The safety of these
substances should be tested by evaluating cytotoxic cell line and compared with commonly used disinfectant benzalkonium-standard (safety).
Project duration:
1-January-2015 - 31-December-2015
Identification of potential protein markers for latency period in non-infections PPROM pregnancies
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
PhamrDr. Juraj Lenčo, Ph.D.; MUDr. Marian Kacerovský, Ph.D.; Mgr. Vojtěch Tambor, Ph.D.
Abstract:
The project aims at characterizing differences in the amniotic fluid protein composition of women with non-infections PPROM with short latency (delivery within 48 hours after rupture of membranes) compared to prolonged latency (delivery after more than seven days following rupture of the membranes).
Project duration:
1-January-2015 - 31-December-2015
Immunomodulators - potential adjuvants based on TLR8 ligands
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Jan Honegr, Ph.D., prof. MUDr. Roman Prymula, CSc., Ph.D., Dávid Maliňák, Ph.D., Rafael Doležal, Ph.D., MSc. Markéta Pasdiorová
Abstract:
The aim of the project is to identify novel potential ligands of TLR8. Compounds will be designed by pharmaceutical chemistry methods utilizing computer modelling - molecullar docking into the active spot of TLR8. Lead compounds from docking study will be synthesized and tested for
cytotoxicity and internal activity on cell line expriming TLR8.
Project duration:
1-January-2015 - 31-December-2015
Metabolic studies of potential anti-Alzheimer’s disease drugs by LC-MS
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Mgr. et Mgr. Rafael Doležal, PhD., PharmDr. Jana Žďárová Karasová, Ph.D., Mgr. Ondřej Soukup, PhD., Mgr. Markéta Pasdiorová, Mgr. Jan Honegr, Ph.D.
Abstract:
One of the most serious neurodegenerative disorders afflicting especially the elderly part of the population is Alzheimer’s disease (AD). This illness manifests itself in vast and irreversible damage of those brain parts ensuring cognitive function like memory, thoughts processing, perception, understanding and oral expressions. Although AD evolves rather gradually, no therapeutics has been found to reverse the pathological neurodegeneration so far. At present, AD patients are treated only with symptomatical drugs (e.g. donepezil, rivastigmine). With the respect that AD global prevalence reached 35 million cases in 2010 and this number is expected to increase geometrically in the future, research of novel anti-AD drugs ranks amongst the most prominent topics in pharmaceutical science.
An integral part of the drug development that possesses a comparatively important position to determination of the pharmacodynamic potential of drug candidates is the analysis of their absorption, distribution, metabolism and elimination characteristics. The aim of this project is to analyze the metabolic properties of selected ten substances with potential effect against AD using human liver microsomal enzymes (HLM). Qualitative and quantitative analyses of the in vitro metabolites will be performed by an ultrahigh-pressure liquid chromatograph and a hybrid mass spectrometer Q Exactive Plus.
Project duration:
1-January-2015 - 31-December-2015
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Jan Korabecny, Dr.; Martin Andrs, Dr.; Petr Jost, Dr.; Ondrej Soukup, Dr.; Kamil Kuca, prof.
Abstract:
DNA-dependent protein kinase (DNA-PK) belongs to the atypical serine/threonine protein kinase family, which play an eminent role in DNA damage response (DDR), especially at double strand breaks. These breaks are
the most dangerous DNA lesions and they are caused mainly by ionizing radiation or several anticancer drugs (e.g., toposisomerase inhibitors). In response to DNA damage DNA-PK and other related kinases trigger complex
signalization network leading to cell cycle arrest, DNA reparation or apoptosis [1,2]. These kinases are important for preserving the genomic integrity; on the other hand they are usually responsible for resistance of tumor cells to DNA-damaging therapy (radiation, various chemotherapeutics). Inhibition of DNA-PK as well as other key proteins involved in DDR brings us a possibility to increase efficiency of standard anticancer therapy [3]. According to newest findings, this way can enable the tumor specific therapy. The majority of tumor cells harbor various mutations and defects in DDR, thus they are more sensitive to inhibition of other components of the network. Therefore, highly selective inhibitors could selectively sensitize these defective tumor cells to cytotoxic cancer therapy [4]. As a template structure for a development of new DNA-PK inhibitors is mostly the non-specific inhibitor of DNA-PK and related kinases 2-morpholino-8-phnylchromen-4-one (LY294002) [5].
[1] Jackson, S. P.;Bartek, J. The DNA-Damage Response in Human Biology and Disease. Nature 2009, 461, 1071–1078.
[2] Hill, R.; Lee, P. W. K. The DNA-Dependent Protein Kinase (DNA-PK): More than Just a Case of Making Ends Meet? Cell Cycle 2010, 9, 3460–3469.
[3] Furgason, J. M.; Bahassi, E. M. Targeting DNA Repair Mechanisms in Cancer. Pharmacol. Ther. 2013, 137, 298–308.
[4] Kaelin, W. G., Jr. The Concept of Synthetic Lethality in the Context of Anticancer Therapy. Nat. Rev. Cancer 2005, 5, 689–698.
[5] Vlahos, C. J.; Matter, W. F.; Hui, K. Y.; Brown, R. F. A Specific Inhibitor of Phosphatidylinositol 3-Kinase, 2-(4-Morpholinyl)-8-Phenyl-4H-1-Benzopyran-4-One (LY294002). J. Biol. Chem. 1994, 269, 5241–5248.
Project duration:
1-January-2015 - 31-December-2015
Development of novel drugs targeted against Alzheimer's disease: combination of cholinesterase inhibitors and antioxidant agents
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
assoc. prof. Daniel Jun; Dr. Jan Korabecny, Dr. Petr Jost, Dr. Eugenie Nepovimova, Dr. Katerina Babkova, Dr. Martina Hrabinova
Abstract:
Alzheimer's disease (AD) is progressive, neurodegenerative disorder affecting mainly people older over 65 age. Apart from neurodegeneration, accumulation of misfolded proteins occurs, e.g. extracellular deposits of amyloid-beta and formation of intracellular tangles of hyperphosphorylated tau protein. These pathological hallmarks represents the mainstream for current drug development process. More in depth, excitotoxicity, mitochondrial dysfunction, neuroinflammation and oxidative stress have been detected as contributing factors to AD pathology. All these events form complex network with oxidative stress playing the crucial role in AD progression and pathogenesis. Reactive oxygen species (ROS) are responsible for neuron impairment in the brains of AD patients. (1) On the contrary, antioxidant species levels are depressed during AD. AD therapy relies on acetylcholinesterase inhibitors (AChEIs), which represents almost the only available drugs mitigating the course of the disease. Current trend in the development of novel AD drugs is focused on so-called "multi-target-directed ligands" (MTDL), agents capable of simultaneous actions toward different pathological mechanisms of the disease. (2) The main aim of this study is to proof the concept of novel MTDLs, which contains a combination of AChEIs with ROS-scavenging moiety.
(1)Rosini, M. et al. J. Med. Chem. 2013, 2821-2831.
(2) Leon, R.; et al. Med. Res. Rev. 2013, 139-189.
Project duration:
1-January-2015 - 31-December-2015
New acetylcholinesterase inhibitors derived from 7-MEOTA structure
Provider:
Internal foundation of University Hospital Hradec Kralové
Investigator:
Martin Mžik, Viktor Voříšek, Vladimír Palička, Jana Žďárová Karasová, Jan Korábečný, Katarina Špilovská
Abstract:
The main aim of this project is evaluation of basic pharmacokinetic parameters (absorption, distribution to central nervous system, elimination and first order metabolites) of centrally acting acetylcholinesterase inhibitors. Two newly synthesized candidates will be choose for this in vivo (male rat, Wistar tribe) evaluation. These new candidates from KS group were chosen after preliminary in silico, in vitro and ex vivo tests.
The new HPLC-MS (high performance liquid chromatography - mass spectrometry) method will be prepared for pharmacokinetic and metabolization study.
Project duration:
1-January-2014 - 31-December-2014
Provider:
Internal foundation of University Hospital Hradec Kralové
Investigator:
Ondřej Soukup, Daniel Jun, Jan Korábečný, Eugenie Nepovímová, Vendula Šepsová
Abstract:
The aim of the project is to prepare approximately 24 triazole, and tetrazole derivatives as potential drugs for Alzheimer's disease, where agonist activity on muscarinic receptors M1 and efficiency in terms of multi-target-directed ligands theory can be expected . Specifically, in vitro agonistic effecr will be determined at M1 receptors, the mechanism will be elucidated and the ability to inhibit acetylcholinesterase will be assessed. Furthermore, an indicative toxicity will be determined by means of cytotoxicity evaluation for the most promising candidates. Another objective is to estabilish a new methodology at the department (PAMPA) assay for determining the ability to cross the blood brain barrier, as an essential prerequisite for drugs acting on the central nervous system.
Project duration:
1-January-2014 - 31-December-2014
Mono-quarternary acetylcholinesterase reactivators as centrally active antidotes against organophosphorus poisoning
Provider:
Internal foundation of University Hospital Hradec Kralové
Investigator:
Jan Korábečný, Ondřej Soukup, Lukáš Werner, Daniel Jun, Rafael Doležal, Veronika Hrabcová
Abstract:
Current therapy of organophosphorus nerve agents and pesticide poisoning combines cholinolytic (atropine), anticonvulsant (diazepam) and acetylcholinesterase (AChE) reactivator based on pyridinium aldoxime (obidoxime, HI-6). However, they are not efficient against the whole spectrum of inhibitors. Due to the presence of quaternary nitrogen, they have low blood-brain barrier (BBB) permeation and thus they are not capable to fully reactivate AChE in the central nervous system (CNS), where they can be responsible for the chronic neural disorders. For this reason, development of novel centrally acting reactivators, that can more efficiently cross BBB is one of the most promising strategy. In this project, we would like to develop two new centrally acting reactivators. This subset will consist of a ligand that will be presumably anchored in the peripheral anionic site of AChE with 3-hydroxypyridine-2-aldoxime moiety responsible for reactivation process.
Project duration:
1-January-2014 - 31-December-2014
Surface active agents as potential disinfectants
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Kamil Musílek, Vladimír Buchta, Daniel Jun, Kamil Kuča, Ondrej Soukup, Marcela Vejsová
Abstract:
The project is aimed to preparation of 20 novel surfactants and evaluation of their effects. The synthesized compounds will be fully characterized (structure and purity determintion) and tested for the antimicrobial activity via various techniques. The selected surfactants with promising antimicrobial activity will be further tested for cytotoxicity effects via standard evaluation. The results of the project should consist in series of surfactants with known antimicrobial and cytotoxic activity, which should determine the perspective for these surfactants in disinfection of surface and skin.
Project duration:
1-January-2014 - 31-December-2014
Proteomics analysis of cerebrospinal fluid in multiple sclerosis patients
Provider:
Internal foundation of University Hospital Hradec Kralové
Investigator:
Martin Vališ, David Doležil, Vojtěch Tambor, Juraj Lenčo, Pavel Šťourač, Pavel Kunc
Abstract:
The goal of this project is to examine the low molecular weight segment of the CSF proteome and compare its composition in the following groups: relapsing multiple sclerosis, clinically isolated syndrome and healthy population. Peptides with altered levels might be considered as potential markers and contribute to the knowledge of the disease pathophysiology. We will employ shotgun proteomic technologies, in order to characterize the low molecular weight CSF proteome.
Project duration:
1-January-2014 - 31-December-2014
Provider:
Internal foundation of University Hospital Hradec Kralové
Investigator:
Martin Vališ, Oldřich Vyšata, Jana Žďárová Karasová, Rafael Doležal
Abstract:
The main aim of this project is established new method for donepezil in biological samples (plasma, cerebrospinal fluid, urine). This highly sensitive HPLC- UV/VIS method should be subsequently used in preclinical and clinical studies. During the method evaluation process, some important parameters will be considered: ease of use, reliability, high sensitivity (under ng/ml) and accessibility in biological /clinical laboratories.
This method will be used in clinical study. The concentration changes will be measured in plasma and cerebrospinal fluid after p. o. administration in different time intervals (pharmacokinetics study). The acetylcholinesterase inhibition potency will be also studied (pharmacokinetic study).
Project duration:
1-January-2014 - 31-December-2014
Diagnostics of adverse effects of a novel syntethised drugs from group of acetylcholinesterase inhibitors; influence to the myoelectric stomach activity in pigs
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Jan Bureš, Marcela Kopáčová, Martin Kuneš, Ilja Tachecí, Jaroslav Květina, Kamil Musílek
Abstract:
Inhibitors of acetylcholinesterase (AChE ) are one of the most effective clinically proven medication in the treatment of cognitive disorders. Indirect parasympathomimetic effect of these agents increases the concentration of acetylcholine at synapses and allows the improvement of cognitive function of the patient. The introduction of a family of AChE inhibitors in clinical practice represents a significant milestone in the treatment of dementia. Currently are used donepezil, rivastigmine and galantamine. However, there are a number of side effects associated with the use of these drugs, including the effects on the gastrointestinal tract, abdominal pain, diarrhea, dyspepsia, indigestion, and more. The use is contraindicated in patients with anamnesis or predisposition to ulcer diseases or in patients with gastrointestinal obstruction.
Project duration:
1-January - 2014 - 1-December-2014
Adjuvants – Immunopotentiators based on TLR Ligands
Provider:
Internal foundation of University Hospital Hradec Kralové
Investigator:
Jan Honegr, Dávid Maliňák, Ondřej Soukup, Rafael Doležal, Kamil Kuča, Roman Prymula
Abstract:
The aim of the project is to identify novel potential ligands of TLR2 and TLR4 with an internal activity comparable with standards - MPL and PamCSK4 (standard for TLR4 and TLR2 respectively) by in vitro high-throughput screening on cell lines stably co-expriming human TLR2 and TLR4. Novel compounds will be designed by of computer assisted molecular modeling with utilization of molecular docking. Hit compounds will be synthesized and tested for cytotoxicity and internal activity. Simultaneously we will be screening compounds from BRC database to found if some of them possess internal activity on TLR.
Project duration:
1-January - 2014 - 1-December-2014
Pharmacokinetics of novel cognitive drugs in the therapy of neurodegenerative disorders
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Martin Kuneš, Jan Květina, Kamil Musílek, Kamil Kuča, Jana Žďárová-Karasová
Abstract:
Acetylcholinesterase (AChE) inhibitors are the most effective clinically proven drug medication in the treatment of cognitive disorders. Alzheimer's disease and others belong into this group of disorders whose are characterised by manifestation of dementia. Part of the pathogenesis of dementia is cholinergic deficit partly due to the inhibition of acetylcholinesterase. Indirect effect of the parasympathomimetic agents increases the concentration of acetylcholine at synapses and allows the improvement of cognitive function of the patient. The introduction of a family of AChE inhibitors in clinical practice is a significant milestone in the treatment of dementia. During the development of new drugs is need to know its pharmacokinetic profiles after administration into the body and to define the therapeutic regimen of the drug.
Project duration:
1-January - 2014 - 1-December-2014
Evaluation of pharmacokinetic properties of novel drug formulations containing platinum cytostatics
Provider:
Internal foundation of University Hospital Hradec Kralove
Investigator:
Martin Kuneš, Jaroslav Květina, Kamil Kuča, Rafael Doležal, Jana Žďárová-Karasová, Michal Pavlík
Abstract:
Colorectal cancers are most widespread types of worldwide cancer disease with high mortality. The therapy with antineoplastics of platinum series are among the most effective therapeutic medication currently available. Their disadvantage is the increasing resistance of tumor cells, systemic toxicity and neuropathy. Present medications consist of repeated intravenous infusion containing the active substance. Recent targeting to the development of oral dosage forms could be from the perspective to the patients, the acceptable dosage and mode of administration, eliminating the adverse effects and direct "delivery" the drug to the site of action, i.e., to the distal parts of the gastrointestinal tract. The development of such dosage form that would meet the requirements defined by a physico- chemical properties of the active substance and also resistance to different pH values in the GIT or microbial flora depends on finding a suitable technological process for preparing such drug formulations.
Diagnostics of adverse effects of a novel syntethised drugs from group of acetylcholinesterase inhibitors; influence to the myoelectric stomach activity in pigs.
Project duration:
1-January - 2014 - 1-December-2014
Non-cholinergic therapeutics for the treatment of Alzheimer's disease
Provider:
University Hospital HK
Investigator:
Ondřej Soukup, Daniel Jun, Kamil Musílek, Jan Korábečný, Ondřej Benek, Vendula Šepsová
Abstract:
The primary objective is to introduce a new methodology to the workplace.Until now, the work was focused on the development of the new drugs in accordance with the cholinergic theory. New targets (BACE1, ABAD and POP) will extend existing know-how, so that it can be used in the the search for new drugs. Then, new methods can be offered in the form of co-operation to other research departments or as service to the private companies.
Another goal is to map non-cholinergic approaches in the treatment of AD and use it in a design of new drugs. Physical output will be cca 20 new chemicals, which will be tested using new methodologies. The most promising candidates will be recommended for the further research or on their stuctural motifs new projects will be submitted.
Project duration:
1-January-2013 - 31-December-2013
Protein and receptor expression analysis in hippocampal adenomas
Provider:
Internal grant agency of the University Hospital Hradec Kralove
Investigator:
Filip Gabalec, Vojtěch Tambor, Juraj Lenčo, Jan Čáp
Abstract:
Hippocampal adenomas account for 10% of intracranial tumors with a prevalence of ~22%. One third of all adenomas do not secrete any hippocampal hormone, thus these are called clinically non-functional adenomas.
In most cases, surgical removal of these adenomas is not complete, which results in frequent recurrence. According to various sources, this can happen in 11-46% of cases. Up to 80% of patients are recurrence-free 5 years after surgery. However, there is no clinically used marker to evaluate the risk of recurrence. The goal of this project is to compare the proteome composition of adenomas from patients with and without recurrence after surgery. We will employ quantitative proteomics method to analyze brain tissue samples.
Project duration:
1-January-2013 - 31-December-2013
Chronic exposition to organophosphate pesticides and its consequences to the changes in brain tissue and association with Alzheimer’s disease
Provider:
Internal grant agency of the University Hospital Hradec Kralove
Investigator:
Daniel Jun, Kamil Kuča, Vojtěch Tambor, Juraj Lenčo, Ondřej Soukup
Abstract:
This project focuses at the hypothesis if a) subchronical exposure to peroral organophosphate pesticide causes observable changes in protein expression in rat brain and b) if these changes are in concordance with general changes observed in neurodegenerative disorders. Based on the obtained results, the project will be potentially expanded to cover both a longer time period and more types of pesticides.
Project duration:
1-January-2013 - 31-December-2013
Development of novel vaccine adjuvants
Provider:
University Hospital HK
Investigator:
Kamil Kuča, Daniel Jun, Roman Prymula, Ondrej Soukup, Pavel Kosina, Jan Honegr
Abstract:
Aim of the project is to establish a novel field of research in University Hospital. We have performed pilot study on novel adjuvants from group of antigen carriers. We have tested 5 novel surfactants´for cytotoxicity. We ave choosed 2 most promising candidates based on cytotoxicity and physico-chemical parameters. These compounds were formulated into the dosage forms wih antigen and most stable and safe formulations are evaluated in-vivo, for ability to enhance the antibody response to the antigen.
Project duration:
1-January 2013 - 31-December-2013
Caffeine derivatives as ATM kinase inhibitors for cancer treatment
Provider:
University Hospital HK
Investigator:
Jan Korábečný, Ondřej Benek, Vendula Šepsová, Ondřej Soukup, Rafael Doležal
Abstract:
Ataxia-telangiectasia mutated (ATM) is serine/threonine protein kinase. ATM simultaneously acts as sensor and enhancer showing the DNA damage, especially double-strand breaks. It is indivisible part of cell cycle regulation during S-phase and take part in G2/M checkpoints. It is also involved in the DNA-reparation processes and indicate DNA-damage as a result of oxidative stress as well. ATM gene mutation leads to hereditary, neurodegenerative disorder so-called ataxia-telangiectasia (A-T, Louis-Bar syndrome). It is characterized by neuropathological syndromes, mainly those parts of brain responsible for coordination and overall body balance, immune system deficits and extreme sensitivity for radiation and DNA-damage inducing agents leading to cancer development. It is illness in population rarely occurring for one in 40000-100000 people. Thus ATM inhibitors can represent useful tool for increase cancer therapy effectiveness and overwhelm resistance to cancer therapy. Caffeine derivatives can be used for sensitization to radiation therapy and to cytotoxic compounds causing DNA damage (topoisomerase inhibitors). The aim of the project is the synthesis novel caffeine derivatives and their in vitro evaluation against ATM.
Duration:
1-January-2013 – 31-December-2013
Provider:
University Hospital HK
Investigator:
Daniel Jun, Kamil Kuča, Vojtěch Tambor, Ondřej Soukup, Juraj Lenčo
Abstract:
The aim of the project is to study the influence of subchronic doses of organophosphorus pesticide on protein expression in rat brain using proteomic methods and to compare the changes in protein expression with the known changes that occur in neurodegenerative diseases.
Duration:
1-January-2013 – 31-December-2013
