Projects / Programmes
Discovering innovative drugs for regulation of haemostasis by venomics of the Vipera a. ammodytes snake
| Code |
Science |
Field |
Subfield |
| 3.06.00 |
Medical sciences |
Cardiovascular system |
|
| Code |
Science |
Field |
| B490 |
Biomedical sciences |
Haematology, extracellular fluids |
| Code |
Science |
Field |
| 3.05 |
Medical and Health Sciences |
Other medical sciences |
Vipera ammodytes ammodytes, venom, haemostasis, anticoagulant phospholipase A2, fibrinolitic metalloproteinase, antitrombotics
Researchers (14)
Organisations (2)
Abstract
Haemostasis is a very subtle, highly regulated system. The precise control of blood coagulation is essential to the life of animals, including humans, so it is no surprise that venomous snakes evolved the arsenal of proteins that target a variety of processes in this system in order to disable their prey or enemy. Some of these molecules can be used to benefit human health. Clinical applications have been found for some of these snake venom proteins, for the majority of them, however, no medical applications currently exist. They represent therefore a vast potential for improving existing or introducing novel therapies and diagnostic procedures in clinics. Nose-horned viper (Vipera ammodytes ammodytes) is a snake whose venom affects in men most severely his haemorrhagic system. The main goal of the proposed project is understanding and exploitment of the venom machinery that produces these effects to discover novel diagnostic and therapeutic procedures for curing human haemostatic disorders. To reach this basic target we will by proteomic approach conclude the mapping of a complete “natural library” of protein/peptide components of the venom. We will identify all components of the venom proteome that interact with the human haemostatic system. Using a range of activity tests we will determine all specific modes of interaction with haemostatic system. On this basis we will group the haemostatically-active components of the venom according to their point of action in haemostatic system and predict their medical applications and/or therapies. We will characterize in detail those haemostatically-active components that will be the most promising according to their pharmacological activity and at the same time by their biochemical characteristics (stability, size, solubility, immunogenicity etc.). For the detailed characterization we will need larger amounts of material therefore we will look for relevant cDNAs in the venom gland library, clone them and prepare recombinant proteins (so called “synthetic library”). For development and optimization we will choose in the first step those medically-interesting molecules that will also by their biochemical features promise straightforward progress towards applications in therapy and/or diagnosis of human haemostatic disorders. In this context, we will give special attention to the low molecular mass components of the venom and the components having no enzymatic activity, for example disintegrins, C-type lectins and three-finger proteins. We will continue the development of the two venom components, ammodytase and ammodytoxin (Atx), which pharmacological activity has already been recognized as medically very interesting. Ammodytase is fibrinolytic metalloproteinase (MP) that will be further developed as a new type antithrombotic lead compound. We will complete its primary structure determination and isolate its cDNA. We will synthesize recombinant ammodytase and, in the process of optimization, some of its domains and their fusions. Recombinant proteins will be pharmacologically tested. Comparing primary structures of the nontoxic MP and homologous hemorrhagic MPs we will define structural basis for hemorrhagicity in this type of enzymes. This will be very important information in designing of ammodytase-based MPs in the process of optimization of MP-based antithrombotics. Atx is a phospholipase A2 with a potent anticoagulant action, which is due to its high-affinity binding to activated blood coagulation factor X (FXa). We will continue with the characterization of interaction between Atx and FXa aiming at designing short peptides or peptidomimetics on the basis of Atx structure interacting with FXa that could be used to prevent blood coagulation. This will include at least in silico molecular modelling of the AtxA-FXa complex. Designed peptides or their analogues will be synthesized and their anticoagulant activity checked. The most promising molecules will be further optimized.
Significance for science
According to World Health Organization (WHO), 29% of global deaths are caused by cardiovascular diseases, qualifying these kind of pathologies as one of the most critical for the mankind. Venous thromboembolic disease is also a major problem in the European Union, where it affects according to the 2007 data more than one million people every year. Moreover, WHO predicts that two leading causes of death in the world in 2030 will be ischemic heart disease and stroke (World Health Statistics 2008, WHO Statistical Information System – WHOSIS). Therefore, one of the strategic priorities of the WHO Cardiovascular Disease Programme (http://www.who.int/cardiovascular_diseases/priorities/en/) is to develop cost effective and equitable health care innovations for management of cardiovascular diseases to reduce morbidity and mortality caused by cardiovascular diseases. Anticoagulants are pivotal for the prevention and treatment of thromboembolic disorders which are one of the most severe medical problems nowadays in developed world indicated by the fact that nearly 1% of the western population receives anticoagulant treatment. It is expected that the problem will expand in the following years, as the population in these parts of the world is rapidly aging. For these reasons, intensive search for novel therapeutic approaches and substances in this medical area is highly relevant.
Significance for the country
The main goal of the proposed project is analysis of the nose-horned venom machinery that affects the human haemostatic system in order to understand and exploit these findings for discovery of novel diagnostic and therapeutic procedures to cure human haemostatic dysfunctions. Expected positive results will have high and immediate impact on improvement of human health. Pharmacologically interesting results will enable further strengthening of our collaboration with either domestic or foreign pharmaceutical industry. Expectedly, the proposed research project will have several indirect positive impacts on our society. These are particularly expected in the field of education. The proposed research activities will in first of all aid to the scientific and professional growth of researchers and university teachers involved – several members of the planned project team are involved in teaching activities at undergraduate and postgraduate levels at different faculties. In the scope of the proposed project students will be educated, a young researcher on her PhD study, as well as several undergraduate students are planned to be involved working on their diplomas. Our knowledge and experiences will be thus directly transferred from the labs to the students. It is our ambition to publish results of our research work in high quality scientific journals and scientific monographs. Together with dissemination of our achievements through lectures on international scientific and medical conferences, we will tend to extend the recognition and scientific reputation of Slovenia in the World. Of the special importance for the Slovenian research sphere and through it for the whole society is also introducing and transferring of the latest research technologies that we are use in our experimental work. Besides proteomics and the surface plasmon resonance technology (SPR), we use in our research also some of the latest functional genomics techniques, such as DNA-microarrays and as the only group in Slovenia the yeast SGA technology.
Most important scientific results
Annual report
2011,
2012,
2013,
final report,
complete report on dLib.si
Most important socioeconomically and culturally relevant results
Annual report
2012,
2013,
final report,
complete report on dLib.si
