Bilateral Project, Modul III, Funded by ANCS, Program PN II ‘Capacities’ Romania - Bulgaria Contract no. 43 / „EVALUATION OF ANTITUMOR AND ANTIVIRAL ACTIVITIES OF NEWLY COORDINATION COMPOUNDS OF BILE ACIDS, SCHIFF AND MANNICH TYPE BASES WITH 3d METAL IONS” Director: Dr. Luminita Patron Lenght:

ROMANIAN PARTNER InstitutionINSTITUTE OF PHYSICAL CHEMISTRY “ILIE MURGULESCU”, ROMANIAN ACADEMY AdressSplaiul Independentei, nr. 202, Bucharest, , Romania Phone/fax/ / / Project leader Dr. Luminita Patron, Head of Coordination and Supramolecular Chemistry Colaborator: Institution INSTITUTE OF CHEMISTRY TIMISOARA AdressTimisoara, strada Mihai Viteazu, nr. 24, Judetul Timis, cod postal Phone/fax/ / / Responsible personDr. Otilia Costisor, Director FOREIGN PARTNER Institution INSTITUTE OF EXPERIMENTAL PATHOLOGY AND PARASITOLOGY, BULGARIAN ACADEMY OF SCIENCE (IEPP - BAS) AdressAcad. Georgi Bonchev Str., Bl. 25, Sofia 1113, Bulgaria Phone/fax/ / / Project leader Dr. Radostina Ivaylova Aleksandrova

INSTITUTE OF PHYSICAL CHEMISTRY “ILIE MURGULESCU” Patron Luminita, PhD Culita Daniela Cristina, PhD Marinescu Gabriela, PhD Beteringhe Adrian, PhD Institute of Chemistry Timisoara Costisor Otilia, PhD Fagadar Cosma Eugenia, PhD Tudose Ramona, PhD Mosoarca Elena Maria, PhD student Vlad Mihaela, PhD student Bucovicean Carmen, PhD student Cretu Daniela – master student Mot Corina – master student Research teams ROMANIAN PARTNER FOREIGN PARTNER INSTITUTE OF EXPERIMENTAL PATHOLOGY AND PARASITOLOGY Alexandrova, Radostina Ivaylova, PhD Alexandrov, Ivaylo Ivanov, DVM, PhD, DSc Alexandrov, Marin Tcviatkov, DVM, PhD Gabrashanska, Margarita Pavlova, DVM, PhD Simeonov, Konstantin Borisov, DVM, PhD Todorova, Iva Tomova Colaborators: Kalfin, Reni Emil, PhD (Institute of Neurobiology, BAS) Leventieva–Necheva, Eleonora Vaniova (Institute of Neurobiology, BAS) Inchovska, Maya Petrova, PhD ((Institute of Experimental Morphology and Anthropology with Museum, BAS) Gluchcheva, Yordanka Georgieva PhD (Institute of Experimental Morphology and Anthropology with Museum, BAS) Shishkov, Stoyan Angelov, PhD (Faculty of Biology, Sofia University “St. Kliment Ohridski” Genova, Petia Dinkova, PhD (National Centre of Infectious and Parasitic Diseases, Sofia, Bulgaria) Timcheva, Constanta Velinova, MD, PhD (National Specialized Hospital for Active treatment in Oncology, Sofia, Bulgaria) – Consultant of the Project MSc Students from Faculty of Biology, Sofia University “St. Kliment Ohridski”

PROJECT DESCRIPTION A general aim of the project presented here is the synthesis of new metal coordination compounds of transition metals with bile acids, Schiff and Mannich bases type ligands and the evaluation of their potential in vitro antitumor activities, as well as their antiviral properties. The knowledge concerning the relationship between the physical and chemical structure of the compounds and their antitumor and antiviral activities in vitro will be obtained. Such structure-activity informations will facilitate the design of drugs with improved anticancer or antiviral properties. The presented project is in accordance with the major priority topic „HEALTH” of the Seventh Framework Programme, aiming to improve the health of European citizens, and with the goals of the „NATIONAL SCIENTIFIC STRATEGIES” of Bulgaria and Romania. The joint work of Bulgarian and Romanian teams on the project will facilitate the exchange of ideas and research data, contacts between scientists of both teams, and will give an opportunity to prepare joint EU project in the future. The aim of the project presented here is the evaluation of cytotoxic and antiproliferative effects as well as antiviral activity of new synthesized coordination compounds of metals (Zn, Cu, Co, Fe, Ni, Pd, V) with bile acids, Schiff and Mannich bases type ligands.

Analysis of the objectives and foreseen benefits This project undoubtedly will also have a favourable educational and practical impact on young scientists, especially PhD and MSc students, post-docs and will influence for the best their research career. The joint work on the project presented here will facilitate the exchange of ideas and research materials, will give opportunities to prepare new EU projects in the future. The proposed objectives are targeting the optimal valorisation of scientific and high-level potential, the consolidation of an excellence Romanian – Bulgarian research network. This project ensures the increase in visibility of the Romanian and Bulgarian science international scale, by publishing the results in ISI indexed journals. The proposed activities join together foundamental and applicative research and large-scale dissemination of the obtained results. The validity and the success chances of the project are revealed by the long-term partnership between the involved parties with the purpose of carrying research activities of great complexity from foundamental to applicative research. Project justification The need to find a safe and highly selective cure for neoplastic diseases remains a major challenge for modern science. The discovery of the antitumor efficacy of cisplatin and some related platinum complexes has stimulated the search for other metals with antineoplastic properties. Different metal complexes, including such of zinc, copper and cobalt, are found to be promising compounds for the design of new anticancer agents. We decided to investigate the antitumor and antivirall properties of such namely compounds because of the following reasons: i)Zinc, copper, cobalt, iron and nickel are essential metals with broad spectrum of biological activities including DNA synthesis (Fe, Co), transport of oxygen, electrons, cell respiration (Fe), normal functioning of the immune and nervous system (Fe, Zn, Co, Cu), etc. ii)There are data about antineoplastic, antimicrobial and antiviral activities of different compounds containing these elements; iii)It was found in our previous investigations that some complexes of Zn(II), Cu(I, II), Co(II), Fe(II, III) and Ni(II) with various ligands (N,N’-bis(4-antipyrylmethyl)-piperazine, N,N’-tetra-(antipyryl-1-methyl)-1,2-diaminoethane, bile acids) expressed cytotoxic and antiproliferative effects on several animal and human tumor cell lines or exhibited antibacterial properties; iv)Numerous biological effects of the ligands have also been reported. The multiple physiological functions of bile acids are well known. Cholic acid, which belongs to bile acid family, and its semisynthetic analogs are reported to inhibit replication of human immunodeficiency virus type 1 (HIV-1) and are also used in the treatment of liver and gastrointestinal disorders. Furthermore, it has been suggested that bile acid conjugates could be helpful in the development of new approaches for targeted anticancer therapy. The cell cycle modulation and induction of apoptosis in several human cancer cells have been reported.

Diagram of project execution Project budget Year 2008: lei Year 2009: lei YearStage description (activities)Partner 2008 Synthesis and physico-chemical characterization of some coordination compounds of transition metal ions containing as ligands anions of bile acids. Romanian Synthesis and physico-chemical characterization of some coordination compounds of transition metal ions containing some Mannich bases as ligands. Romanian Evaluation of cytotoxic/antiproliferative and antiviral activities of the synthesized during the 1 st year coordination compounds in cell cultures. Bulgarian 2009 Synthesis and physico-chemical characterization of some coordination compounds of transition metal ions containing as ligands Schiff bases and anions of bile acids. Romanian Synthesis and physico-chemical characterization of some coordination compounds of transition metal ions containing Mannich bases as ligands. Romanian Evaluation of cytotoxic/antiproliferative and antiviral activities of the synthesized during the 2 nd year coordination compounds in cell cultures. Investigations on the in vivo antitumor effect of the most promissing compound(s). Bulgarian

Results: 1.“Cobalt and cobalt compounds – biological activity and potential antitumor properties”, Radostina Alexandrova, Reni. Kalfin, Otilia Costisor, Luminita Patron, et.all., Proceedings of The second Edition of the Symposium “New trends and strategies in the chemistry of advanced materials with relevance in biological systems, technique and environmental protection”, 6-7 november 2008 Timisoara, Romania, pag “The antimicrobial power of metals and metal compounds”, Radostina Alexandrova, Ramona Tudose, Elena Mosoarca, Daniela Culita, Gabriela Marinescu, Proceedings of The second Edition of the Symposium “New trends and strategies in the chemistry of advanced materials with relevance in biological systems, technique and environmental protection”, 6-7 november 2008 Timisoara, Romania, pag “New heteropolinuclear complexes of Cu(II) and Zn(II) containing a pyrazolone derivative as ligand. Synthesis and spectral characterization”, Elena Mosoarca, Ramona Tudose, Otilia Costisor, Proceedings of The second Edition of the Symposium “New trends and strategies in the chemistry of advanced materials with relevance in biological systems, technique and environmental protection”, 6-7 november 2008 Timisoara, Romania, pag “Synthesis and characterization of 3d complexes with N,N’-bis(antipyryl-4-methyl)piperazine”, Ramona Tudose, Elena Mosoarca, Otilia Costisor, Proceedings of The second Edition of the Symposium “New trends and strategies in the chemistry of advanced materials with relevance in biological systems, technique and environmental protection”, 6-7 november 2008 Timisoara, Romania, pag “Eefectul combinatiilor complexe ale Co(II) cu ligand de tip baze Mannich asupra diviziunii mitotoce la Alium Cepa L”, Ramona Tudose, Elena Mosoarca, Irina Petrescu, Otilia Costisor, Proceedings of The second Edition of the Symposium “New trends and strategies in the chemistry of advanced materials with relevance in biological systems, technique and environmental protection”, 6-7 november 2008 Timisoara, Romania, pag “Coordination compounds of Co(II), Ni(II), Cu(II) and Zn(II) with ursodeoxycholic acid”, Daniela C. CULIŢĂ, Gabriela MARINESCU, Luminiţa PATRON, Floriana ŢUNĂ, Nicolae STANICĂ and Ma YUN, Revue Roumaine de Chimie, in press, 2009.

7. “Spectral characterization and biological activity of Cu(II) complex containing N,N’-tetra-(4-antipyrylmethyl)-1,2 diaminoethane (TAMEN) as ligand”, Elena Maria Mosoarca, Ramona Tudose, Livia Avram, Mihai Milos, Carmen Bucovicean, Carmen Cretu, Corina Mot, Viorel Sasca, Radostina Alexandrova, Otilia Costisor, Proceedings of The Third Edition of the Symposium With International Participation New Trends and Strategies in the Chemistry of Advanced Materials, November 5-6, 2009 Timisoara, Romania, p “Copper Compounds as Possible Antitumor Agents”, R. Alexandrova, R. Kalfin, E. Leventieva-Necheva, E.M. Mosoarca, R. Tudose, O. Costisor, D.C. Culita, G. Marinescu, L. Patron, Proceedings of The Third Edition of the Symposium With International Participation New Trends and Strategies in the Chemistry of Advanced Materials, November 5-6, 2009 Timisoara, Romania, p Scientific Communications at National and International Scientific Meetings