First year of the project

Much emphasis has been paid on characterization of the role of selected protein kinases in signal transduction pathways and in various diseases models such as proliferation on tumour cells, functions of immune cells and vascular cells, including endothelial and smooth muscle cells and also in central nervous system physiology. Part of the results from these studies have been published in several scientific publications.

In a relatively short period of time several hundred natural extracts have been prepared, pure compounds isolated and preliminary activity screening has been done. Also several hundred synthetic compounds targeted to various protein kinases have been synthesised.

A number of the novel compounds show biological activity in various target-based assay systems utilising purified kinase enzyme preparations. In comparison with pure synthetic compounds some of the purified extracts have been more efficacious in cell culture (functional-based assay). Several protein kinase inhibitors showed inhibitory activity in vascular smooth muscle proliferation assay and therefore it is possible that in the future protein kinase inhibitors may be used in the treatment of re-stenosis after balloon surgery (angioplasty) of coronary arteries.

More than 100 compounds originating from consortium laboratories have been tested in anti-parasite assays against Leishmania species. Some compounds show promising activity and this observation can open new avenues in the treatment strategies of parasite diseases in the future. Natural and synthetic compounds have been tested in models of leukaemia and already two plant-derived compounds and one synthetic PKC-modulating compound show high potency against leukaemic cells. The effects of selected inhibitors on neuronal excitotoxicity (models of neuronal death identified during stroke) were tested and identified as neuroprotective agents in models of stroke. One protein kinase inhibitor has inhibited neuronal excitability in an in vitro model of epilepsy. The lead compound and its successors will be further studied for possible future antiepileptic uses.

An improved method for early characterisation of cell permeation properties of newly discovered compounds is in development. This method will be automated and fully developed during the project lifetime and will give early estimate of “drugability” of novel compounds. Anaplastic Lymphoma Kinase (ALK) is aberrantly expressed and activated in certain cancers. An enzyme-linked immunosorbent assay (ELISA) for measuring ALK kinase activity has been developed which enables higher throughput screening of new compounds affecting ALK-kinase. In this assay as well as in many other kinase assays large quantities of kinase proteins are needed and several laboratories in the consortium are producing multiple recombinant proteins for assay purposes.

Rational drug design is informed by three dimensional structures and models of proteins and more specifically of the active site – usually the ligand binding site is used for discovering novel chemical entities and optimization of lead compounds. Several laboratories in the consortium are producing in various cell cultures selected protein kinases in large quantities in order to determine crystal structure of the kinases in question. Crystallisation trials for the first of these are in progress.

Expected impact

The research groups of the Pro-KinaseResearch consortium represent different disciplines of science with the same focus of interest namely the protein kinases. Our research efforts aim at the discovery of new means by which the activity of protein kinases can be modified. Our research will increase the possibility for development of clinically useful drugs having a novel mechanism of action.

In Europe and other areas of the World with a high standard of living, diseases such as cancer, autoimmune diseases, cardiovascular diseases and neurological diseases are an increasing threat to public health and a drain on public resources. In developing countries, parasitic infectious diseases such as malaria and leishmania are enormous health problems. In the future, innovations based on our results can be made available to a global drug market to combat the diseases mentioned above.