Development of novel acetylcholinesterase inhibitors as treatment of Myasthenia Gravis
Musílek, K.
Supported by the Czech Republic Grant Agency, 2009–2011 (Project No.: GP203/09/P130)
The treatment of autoimmune disease Myasthenia gravis (MG) is based on a combination of various drugs. The acetylcholinesterase (AChE) inhibitors are used at the first stage of MG (e.g. pyridostigmine bromide) as a symptomatic treatment. These drugs are effective both in peripheral and central nervous system (cross blood-brain barrier – BBB). Their administration causes a huge amount of undesirable effects and it is necessary to reduce their dosage. The preparation, in vitro testing and structure-activity relationship (SAR) of novel AChE inhibitors will be done in our project. Novel compounds will be designed as the bispyridinium molecules with various functional groups and linkers. The bisquaternary structure should guarantee the minimal penetration through BBB and their peripheral effectiveness. SAR will be considered via experimental results (in vitro – IC50) and quantitatively via computer programmes (QSAR). The peripheral AChE inhibitors suitable for application in vivo on animals and for further testing as relevant candidates for MG treatment should become the project results.
Development of novel antidotal treatment against organophosphorus pesticides
Musílek, K., Kuča, K., Pohanka, M., Žďárová Karasová, J., Marek, J.
Supported by the Ministry of Education, Youth and Sports, 2009–2012 (Project No.: ME09086)
The development of the broad-spectrum acetylcholinesterase reactivator against organophosphorus pesticides (OPP) is the main aim of the whole project. The synthesis and in vitro evaluation of novel or formerly prepared compounds will be used for this purpose. The structure-activity studies will be figured out for its determination. Such reactivator will be further tested in vivo and might become a candidate for the preclinical trials against OPP.
FARMAKO – Determination of important pharmacokinetic and biochemical parameters and evaluation of blood-brain barrier penetration using drugs introduced to Czech Army
Žďárová Karasová, J., Pohanka, M., Novotný, L., Kuča, K.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200811)
The aim of this project is to characterize pharmacokinetics of substances using in therapy by intoxications. Mainly their distribution in body tissues, in vitro and in vivo testing of ability to penetrate into central nervous system and their possibility to injury brain. At the same time we will evaluate some important biochemical data, which can be influenced by this therapy.
INDIKÁTORY II – Reverse detection of received ionizing radiation dose by monitoring of cell population changes using biophysical methods
Šinkorová, Z., Tóthová, I., Pejchal, J., Zárybnická, L., Kročová, Z., Österreicher, J.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200809)
To establish extrapolated calibration curve for determination the received dose in individuals exposed to ionizing radiation which could be practical used in the Czech army and to design the best enlistment of this biophysical method into the therapeutic-transfer system of the Czech army.
INHIBITOR – Novel inhibitors of acetylcholinesterase as prophylaxis of nerve agent poisonings
Musílek, K., Žďárová Karasová, J., Pohanka, M.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200805)
This project is focused on finding of suitable reversible acetylcholinesterase inhibitor as a replacement of prophylactic drug pyridostigmine. For this purpose, 100 reversible AChE inhibitors will be prepared, evaluated in vitro and compared with prophylactic drugs used in the Army of the Czech Republic. Regarding the structure-activity relationship, the best reversible AChE inhibitor will be determined and recommended for in vivo evaluation.
MORČE – Influence of the nerve agent and reactivators of acetylcholine esterase on the Guinea pig
Novotný, L., Žďárová Karasová, J., Kuča, K., Pohanka, M., Jun, D.
Supported by the Czech Republic Ministry of Defence, 2009–2011 (Project No.: OVUOFVZ200905)
The aim of this project is determine the physiological activity of Acetyl Choline Esterase (AChE) and detection of LD50 of inhibitors paraoxon, tabun, cyklosarine and reactivators AChE pralidoxim, obidoxim, trimedoxim, HI-6, methoxim. Next task is asses protective effectively of reactivators. Also detection of AChE activity in organs after intoxication with nerve compound and following reactivation. We will also follow up plasmatic concentration of reactivatos in plasma after i.m. administration and morphological changes in the tissues.
ORCHIDS – Evaluation, optimisation, trialling and modelling procedures for mass casualty
Simpson, J., Kuča, K., Cabal, J.
Supported by the Executive Agency for Health and Consumers, 2008–2011 (Project No.: 100940)
The ORCHIDS project (Optimisation through Research of Chemical Incident Decontamination Systems) involves the evaluation of emergency decontamination methods, and the exercising and modelling of established mass casualty decontamination facilities. The project will deliver quantitative evidence on the optimum techniques for dealing with a range of potential contaminants and scenarios requiring emergency decontamination. A full range of issues will be addressed, from applied toxicological research to mass casualty decontamination exercising and modelling. This applied research will generate evidence-based guidelines on the optimum techniques for effective mass casualty decontamination, which will be disseminated by the project team through a network of EU Partners and stakeholders, which will be established and developed during the project. In addition, the project will consider the provision for minority and vulnerable groups in emergency decontamination, and will produce public information materials (leaflets and educational tools) designed to help increase knowledge, trust and confidence in emergency decontamination provision.
OTRAVA – Novel prophylactic antidotes of nerve agent poisonings based on scavengers
Jun, D., Musílek, K., Pohanka, M., Kuča, K.
Supported by the Czech Republic Ministry of Defence, 2009–2012 (Project No.: OVUOFVZ200902)
Organophosphorus compounds are weaponized in some countries as chemical warfare agents (sarin, tabun, VX). These compounds inhibit enzymes acetylcholinesterase and butyrylcholinesterase. As antidotes of poisonings by these compounds are often used anticholinergics (atropine, benactyzine) and cholinesterase reactivators (pralidoxime, obidoxime, HI-6). Favorable solution is using of specific enzymes (mostly cholinesterases) as scavengers, able to catch toxic organophosphorus compounds in the bloodstream before they start their toxic effect in the organism. We would like to find suitable cholinesterase reactivators, with the aim to suggest and test their combination with enzymes as pseudocatalytic bioscavenger. This combination allows increase prophylactic efficacy of administered enzyme.
PROTEIN – Biosensors for determination of nerve agents and yperites using recombinant proteins and nanotechnology
Pohanka, M., Žďárová Karasová, J., Musílek, K., Kassa, J., Kuča, K.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200807)
>The proposed project is aimed at construction of enzyme based biosensors with immobilized ether cholinesterase and/or dehalogenase for detection of nerve agents and yperite using highly innovative nanotechnologies.
REAKTIVÁTOR – Robotized system for in vitro evaluation of novel rectivators of acetylcholinesterase inhibited by nerve agents
Cabal, J., Pohanka, M., Musílek, K., Kuča, K., Jun, D.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200801)
The aim of this project is to develop new robotic system based on sequential injection analysis (SIA) for testing of reactivation potency of newly synthesized antidotes against nerve agent poisonings.
RONSDOZ – Noninvasive measurement of proinflamatory markers of oxidative stress in irradiated as an indicator of received dose of radiation. Protective role of acetyl-L-carnitine
Vávrová, J., Řezáčová, M., Pejchal, J., Österreicher, J., Tichý, A.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200806)
To evaluate whether the bioindicators of oxidative and nitrative stress and inflammations of the airways can be used as biodosimetric markers after exposure to gamma radiation, after whole-body and partial irradiation of rats. To evaluate protective effect of acetyl-L-carnitine.
SUBSTANCE – Development of novel decontaminants and disinfectants of skin based on micellar compounds
Kuča, K., Novotný, L., Musílek, K., Pohanka, M., Jun, D., Cabal, J., Kassa, J.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200803)
The aim of this project is a development of new compound or mixture with a good decontaminant efficacy against wide spectrum of chemical warfare or pesticides. The second aim will be also to gain the universal disinfectant, which will be effective against many kinds of microbes.
The evaluation of potency acetylcholin-esterase reactivators to penetrate through the blood-brain barrier
Žďárová Karasová, J., Kuča, K., Novotný, L.
Supported by the Internal Grant Agency of the Czech Republic Health Service, 2009–2010 (Project No.: NS9748)
A penetration of different acetylcho-linesterase reactivators through the blood-brain barrier will be studied. Firstly, the AChE reactivators will be assessed be means of a novel analytical method in vitro. Based on previous results, the promising reactivators for in vivo evaluation will be chosen on rat animal model (tribe Wistar). Finally, the time-course changes in levels of reactivators in plasma and central nervous system will be determined.
The role of virus associated cellular proteins in T-lymphocyte dysfunction
Boštík, P., Hubálek, M., Řehulka, P., Červený, L., Pejchal, J., Kročová, Z., Boštíková, V.
Supported by the Czech Republic Grant Agency, 2010–2014 (Project No.: GAP304/10/1161)
Herpetic viruses, such as VZV, and lentiviruses, such as HIV or SIV, are enveloped viruses, which infect CD4 T cells and cause transient (VZV) or progressive (SIV) dysregulation of T cell function. This effect is mainly indirect, as the fraction of infected cells is small, but the dysregulatory effect is observed in much larger cell population. These viruses incorporate host-derived proteins into their envelopes during the process of virus maturation and these proteins can either retain their function or engage their receptors and subsequently initiate intracellular signaling. This can be mediated by Akt-GSK3 pathway and PGE metabolism, leading to T cell dysfunction and apoptosis. This proposal utilizes state-of the art proteomic approach to Identification of host cell proteins incorporated into the SIV and VZV virions. The role of these host cell proteins will be subsequently investigated in their effects on CD4 T cell signaling cascades and can therefore lead to the elucidation of mechanisms involved in CD4 T cell dysfunction and death in such diseases as chickenpox and AIDS.
YPERIT – Potential interference of toxic properties of sulphur mustard
Štětina, R., Svobodová, H.
Supported by the Czech Republic Ministry of Defence, 2008–2011 (Project No.: OVUOFVZ200810)
The aim of this project is to estimate: 1)The influence of poly-(ADP ribose) polymerase (PARP) inhibitors on to the cytotoxic and cytostatic effect of sulphur mustard . The influence of of PARP inhibitors on the DNA repair (repair of cross-links) induced by SM will be studied in detail in mammalian and human cells. Both normal and DNA repair deficient mutant cells will be used. 2) The influence of selected antioxidants on the cytostatic effect of SM. The oxidative DNA damage and its relationship to the cytotoxicity will be studied. 3) The influence of combined treatment with PARP inhibitors and antioxidants on the cytostatic effect of SM.