Metallo – Porphyrin Compounds As Anti
Total Page:16
File Type:pdf, Size:1020Kb
Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2012 Metallo – Porphyrin Compounds as Anti-Cancer Compounds and their Interaction with Different conformations of DNA: Syntheses, Model and in vitro Studies of Metallo – Porphyrin Compounds Antoni, P Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-60422 Dissertation Published Version Originally published at: Antoni, P. Metallo – Porphyrin Compounds as Anti-Cancer Compounds and their Interaction with Dif- ferent conformations of DNA: Syntheses, Model and in vitro Studies of Metallo – Porphyrin Compounds. 2012, University of Zurich, Faculty of Science. Metallo – Porphyrin Compounds as Anti- Cancer Compounds and their Interaction with Different Conformations of DNA Syntheses, Model and in vitro Studies of Metallo – Porphyrin Compounds DISSERTATION zur Erlangung der naturwissenschaftlichen Doktorwürde (Dr. sc. nat) vorgelegt der Mathematisch – naturwissenschaftlichen Fakultät der UNIVERSITÄT ZÜRICH von Philipp M. Antoni aus Deutschland Prüfungskomitee Prof. Dr. Roger Alberto (Vorsitz) Prof. Dr. Roland K. O. Sigel (Beisitz) PD Dr. Bernhard Spingler (Leitung der Dissertation) Zürich 2012 1 Metallo – Porphyrin Compounds as Anti- Cancer Compounds and their Interaction with Different Conformations of DNA Syntheses, Model and In Vitro Studies of Metallo – Porphyrin Compounds 1. List of Synthesized Compounds _______________________________________ 5 2. Acknowledgements _______________________________________________ 11 3. Abstract ________________________________________________________ 14 3.1 Deutsche Zusammenfassung ____________________________________ 14 3.2 English Abstract ______________________________________________ 16 4. Interaction of Metallo – Porphyrins with Different Conformations of DNA ___ 18 4.1 DNA – an introduction _________________________________________ 18 4.1.1 The early days of DNA research ____________________________________________ 18 4.1.2 DNA and its role as a target of molecular chemistry ____________________________ 22 4.1.3 Different conformations of DNA ____________________________________________ 25 4.1.4 Formation of Z-DNA _____________________________________________________ 31 4.1.5 Circular dichroism[80] _____________________________________________________ 34 4.1.6 Possible biological functions of Z-DNA. ____________________________________________ 36 4.2 Abilities of Porphyrins to detect DNA _____________________________ 38 4.2.1 Interactions with Z - DNA _________________________________________________ 38 4.2.2 Interaction of porphyrins with G – Quadruplexes ______________________________ 40 4.3 Background and motivation for this work _________________________ 41 4.4 Results and Discussion – Interaction of Porphyrins with DNA __________ 42 4.4.1 Interaction of metallo – porphyrins with different conformations of DNA ___________ 42 4.4.2 Interaction of lead porphyrins with guanosine_________________________________ 43 4.4.3 Interactions of porphyrin treated poly d(GC) with Z- inducing agent _______________ 48 2 4.4.4 Interactions of lead porphyrins with previously induced Z- conformation of poly d(GC) 51 4.4.5 Calculations of the coordination of zinc porphyrin towards DNA __________________ 55 4.4.6 Z-DNA detecting molecules _______________________________________________ 59 4.4.7 Porphyrins as side arms: an odyssey ________________________________________ 63 4.4.8 Corner Building Blocks ___________________________________________________ 66 4.4.9 Synthesis of the Bridges __________________________________________________ 67 4.5 Conclusion and Outlook ________________________________________ 71 5. Photoactive Metalloporphyrins for a Combined Chemo- and Photodynamic Therapy ____________________________________________________________ 73 5.1 Abstract _____________________________________________________ 73 5.2 Introduction _________________________________________________ 74 5.2.1 Cisplatin _______________________________________________________________ 74 5.2.2 Second and Third Generations of cisplatin analogues ___________________________ 75 5.2.3 Photodynamic Therapy with Porphyrins[197, 276-280] ______________________________ 80 5.2.4 Mechanism of Porphyrin Excitation _________________________________________ 82 5.2.5 Porphyrins and their Biological Behavior _____________________________________ 85 5.2.6 Approved Drugs containing Porphyrins ______________________________________ 86 5.2.7 Combinatorial Therapies __________________________________________________ 88 5.3 Motivation and objective of this project___________________________ 90 5.4 Results and Discussion _________________________________________ 92 5.4.1 Synthesis and Characterization _____________________________________________ 92 5.4.2 Stability of Platinum-Porphyrin Complexes in Water and Serum ___________________ 98 5.4.3 Porphyrin Complexes and their interaction with DNA __________________________ 103 5.4.4 Cytotoxicity Studies in Carcinoma Cells _____________________________________ 107 5.4.5 Calculated Interactions __________________________________________________ 117 5.5 Conclusion and Outlook _______________________________________ 120 6. Conclusion and outlook ___________________________________________ 123 7. Experimental ___________________________________________________ 126 7.1 List of Abbreviations _________________________________________ 126 3 7.2 Instrumentation and materials _________________________________ 129 7.3 Synthesis of Porphyrin and their Building Blocks ___________________ 132 7.4 Synthesis of Metallo - Porphyrin Complexes ______________________ 149 7.5 Organic Bridges ______________________________________________ 155 7.6 Platinum building blocks ______________________________________ 161 7.7 Multi – porphyrin and Platinum Arrays___________________________ 163 7.8 CD titration of poly (dG-dC) - poly (dG-dC) with Cu2+, Ni2+ and Zn2+ porphyrins _______________________________________________________ 168 7.8.1 UV, Fluorescence, and CD titration of 5’-GMP with (metallo)-porphyrins ___________ 168 7.8.2 UV, Fluorescence, and CD titration of poly d(GC) with (metallo)-porphyrins[379] ______ 168 7.8.3 HPLC Measurements[343] _________________________________________________ 168 7.9 MTT Cytotoxicity Studies ______________________________________ 169 7.9.1 Cell Lines [380, 381] _______________________________________________________ 169 7.9.2 MTT Cytotoxicity assay [38, 347, 382, 383] ________________________________________ 169 7.9.3 Crystallographic Data ___________________________________________________ 170 8. Literature ______________________________________________________ 173 4 1. List of Synthesized Compounds 5 6 7 8 9 10 2. Acknowledgements I would like to thank colleagues, friends and family that supported me during the last 4 years. It has been an intense and interesting time at the University with a lot of new and exciting experiences, tasks, ups & downs and friendship. A lot of persons helped me in various aspects, personally and professionally. In the need of help, I could rely on a precise and fair feedback, support and also some open ears. During the time at the University of Zurich, I received a lot of good ideas which made up my mind how I want to form my future. Lots of persons joined in this part of my life and accompanied me for a while. I would like to thank all those for their great support. Especially the friendly help, constructive advices, ideas and patience of the members of the Inorganic Institute of the University of Zurich was very important for this work. Thank you! I would also like to thank my friends for their persistence, patience and support. I am very happy and grateful that you supported me. In particular, I would like to acknowledge the following persons… PD Dr. Bernhard Spingler, for your patience, advice, and support. Thank you for giving me this interesting topic to work on and free hands to develop it further. Thank you for teaching me how to develop new ideas, working techniques and open thoughts in various topics, chemically and personally. Thanks for your friendship and the many discussions and for the correction of this supervision and work. Prof. Dr. Roger Alberto, for giving me the opportunity to work in his group and giving me the freedom to develop and experiment on many ideas. I enjoyed a lot organizing our yearly Christmas lecture – it was a lot of fun. I enjoyed the Trin/Kandersteg meetings and the open atmosphere in your group. I appreciated a lot your direct way of communication, especially during the writing of my thesis. Thank you for helping me keeping my focus on my thesis and for your supervision. 11 Prof. Dr. Roland Sigel, for helping me in different aspects during my time as a PhD student. We built the CMSZH and had controversial discussion and common battles in various commissions. Thank you for helping me developing my profile and the many good discussions. Prof. Dr. Eva Freisinger, for her support, patience, open doors for discussions. I always appreciated the time we discussed our various topics. Thanks. My labmates and students: Stefan Schnidrig, for a great time in the lab synthesizing a lot of challenging molecules and combining them to functionality. Michael Felber and Miriam Oberholzer for their highly motivated, skilled work and support. Dr. Benji Probst, for a great time and support from Vordiplom to PhD, lot of exchange of experience and some memorable