“SEE-PROT” & “SEE-PHARM” are the protein production and the preclinical evaluation of molecules, modules of SEE-DRUG project. The tools offered for these efforts are protein production and characterization facilities an upgraded confocal microscope, an intravital microscope and myographs.

SEE-PROT: Production of protein & peptides and screening/optimization of polypeptide samples, for structural and functional studies either by NMR or by X-ray and other biophysical techniques. This module will allow UPAT’s researchers and researchers from other regional (Greek, South-Eastern EU or other) institutes, to gain experience, to collaborate with SEE-DRUG experts and finally to achieve the identification, cloning, production and characterization of new protein drug-targets that will be forwarded for structural characterization, and for the study of their interaction with other protein/peptide partners and drug screening. The availability of proteins suitable for structural analysis constitutes a major bottleneck in structural biology and structure-based drug design methodologies. Although over-expression in bacteria is a well-established process and the collaborating groups of the SEE-DRUG consortium have a long-term experience in the efficient production of peptide and protein samples for structural and functional studies, there are certain types of proteins, involved in biochemical pathways of tumour growth and/or suppression, cardiovascular and neurodegenerative diseases that are the main research targets of the team, and which may need the incorporation of eukaryotic or mammalian expression systems. Additionally, a state-of-the-art NMR methodology relies on the investigation of proteins, RNA and DNA that are uniformly or selectively enriched with NMR-active nuclei, such as 13C and 15N, while sometimes requires combination of the enrichment to the above nuclei with 1H substitution by 2H nuclei.

SEE-PHARM: Disease-modelling cell-based assays including live cell imaging techniques, complemented by ex vivo and in vivo screening, to test protein-protein/peptide and protein-drug lead interactions, evaluate efficacy of candidate molecules, determine new biomolecular interactions and study the effects of the leads on basic cellular pathways. SEE-PHARM’s role will therefore be to use all its available tools to test bio-molecules developed on NMR structure-based information and provide valuable feedback regarding their optimization. The upgrading of the existing Confocal Microscopy Facility, will comprise, according to our knowledge, a unique facility in Greece for its capacities, properties and applications. Additionally, the UPAT groups of Molecular Pharmacology has adequate state-of-the-art instrumentation to evaluate a broad range of processes in vitro (e.g. protein and mRNA analysis, ELISA, luciferase and fluorescence plate readers etc), being able to functionally evaluate in vivo or ex vivo some critical processes in cardiovascular and inflammatory diseases. The Intravital microscope and the Myographs: a) will significantly upgrade the groups’ ability to move lead compounds to a more significant level and b) will be perfectly complementary to the existing infrastructure. The ability to perform this testing can support the application and concur to the development of more solid intellectual property, eliminate a compound as ineffective or indicate pharmacokinetic and metabolism-related properties that can be improved and optimized. For this reason we request to add the following two setups that allow us to go a step further in our drug candidate characterization-oriented effort, which will also considerably enhanced by the information gained by NMR and Xray structural studies.