Natural Products with anticancer activity from Moroccan plant Thymelaea lythroides and its endophyte Chaetomium aureum Dissertation zur Erlangung des akademischen Grades Doktor der Naturwissenschaften (Dr. rer. nat.) und Doktor der Biowissenschaften und der Medizin (Dr.) vorgelegt der Bergischen Universität Wuppertal Fachbereich C – Mathematik und Naturwissenschaften und Mohammed V-Souissi Universität Rabat Fakultät – Medizin und Pharmazie von Fatima Zahra Kabbaj Wuppertal 2013 Aus der Arbeitsgruppe der Organischen Chemie der Bergischen Universität Wuppertal Diese Dissertation kann wie folgt zitiert werden: urn:nbn:de:hbz:468-20140404-113126-9 [http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:468-20140404-113126-9] Gedruckt mit der Genehmigung der Mathematisch-Naturwissenschaftlichen Fachbereich C der Bergischen Universität Wuppertal und Medizinisch-Pharmazeutischen Fakultät der Mohammed V-Souissi Universität Rabat Referent: Prof. Dr. Hans-Josef Altenbach Koreferent: Prof. Dr. Moulay El Abbes Faouzi Tag der mündlichen Prüfung: 25.11.2013 Erklärung Hiermit erkläre ich ehrenwörtlich, dass ich die vorliegende Dissertation mit dem Titel „Natural Products with anticancer activity from Moroccan plant Thymelaea lythroides and its endophyte Chaetomium aureum“ selbst angefertigt habe. Außer den angegebenen Quellen und Hilfsmitteln wurden keine weiteren verwendet. Diese Dissertation wurde weder in gleicher noch in abgewandelter Form in einem anderen Prüfungsverfahren vorgelegt. Weiterhin erkläre ich, dass ich früher weder akademische Grade erworben habe, noch dies versucht habe. Wuppertal, den 21.10.2013 Fatima Zahra Kabbaj I dedicated this work to my husband and my parents Acknowledgement Acknowledgement First and foremost thanks to the Almighty God “ALLAH” who has granted me all these graces to fulfill this work and blessed me by His power, mercy and patience during my life. To Him I extent my heartfelt thanks. It is a pleasure to find the chance to thank all the people that were directly or indirectly involved in the success of the completion of this doctoral thesis. First of all, I would like to acknowledge Prof. Dr. rer. nat. Hans-Josef Altenbach and Prof. Dr. Moulay El Abbes Faouzi for the excellent working environment, for their patience and encouragement during my first steps in the field of natural product research, and for taking over the supervision of my work with their steady support until the completion of this thesis. I would like to express my cordial thanks and gratitude to Prof. Dr. rer. nat. Peter Proksch for giving me the opportunity to pursue my doctoral research at his institute, as well as for his valuable recommendations, his fruitful discussions, his unlimited support and for the excellent working facilities at the Institute of Pharmacological Biology and Biotechnology, Heinrich- Heine-Universität, Düsseldorf. My gratitude and all my regards to Dr. Abdessamad Debbab for his instructive supervision, his kind help, his continuous support and encouragement throughout the completion of this work. Without his guidance this thesis would not be what it is today. My special thanks to Dr. Aly Hassan Amal and Dr. Lai Daowan for their constructive advises, sharing their expertise in NMR data interpretation as well as for their help and support in good and bad times. I would like to deeply thank Prof. Yahia Cherrah (Chief of the Division of the National Laboratory of Control of Drugs in Morocco) for his guidance, his continuous encouragement, and his kind advices. ii Acknowledgement I am deeply grateful to all members of the jury for agreeing to read the manuscript and to participate in the defense of this thesis. I would like to express my profound gratitude to my professors and colleagues from the Department of Pharmacology, University Mohammed V, Rabat, Morocco, especially Prof. Abdelaziz Bouklouze for defending me during the selection of the candidates of CeDoc at the Faculty of Medicine and Pharmacology of Rabat in Morocco. My gratitude and all my regards to Prof. Jamal Taoufik (Director of the Doctoral School “Sciences de la Vie et de la Santé” and Vice Doyen of Pharmacy at the Faculty of Medicine and Pharmacy) for giving me the opportunity to access the Doctoral School “Sciences de la Vie et de la Santé” and for carrying out the success of this first education in Morocco. My sincere gratitude and thankfulness to the professors of ongoing seminars at the Doctoral School “Sciences de la Vie et de la Santé”; for their coaching, assiduity and their sense of responsibility. I would like to thank Prof. Maati Nejmi (Director of the NIO) who allowed me to carry out the ethnpharmacological survey at the National Institute of Oncology in Rabat. My special thanks also for Dr Abdelouahad Erraki (Head register of the NIO) for his valuable help to get the information concerning the questioned patients. I would like also to thank the clinicians and staff of chemotherapy and radiotherapy for their collaboration. Not to forget to extend my thanks to the patients of NIO for their cooperation. My special thanks to Prof. Abdelaziz Benjouad (Vice President of Research and Development at the International University of Rabat), who guided me patiently in my first year and help me to select the natural source of this study. Many thanks for the friendly cooperation to Prof. Dr. Ahmed Chadli, at the Molecular Chaperone/Radiobiology & Cancer Virology in Georgia Regents University, for carrying out the cytotoxic inhibitors of Hsp90 tests, and for his fruitful discussions by interpreting the results. I would like to specially thank the people who are involved with my success as following; iii Acknowledgement Dr. Victor Wray (Gesellschaft für Biotechnologische Forschung, Braunschweig) for aiding the chemical structure elucidation. Prof. Dr. Wen Han Lin (Natural Research Laboratory of Natural and Biomimetic drug, Baijing Medical University, Beijing, Republic of China) for the suggestion of structure elucidation of compounds isolated from fungus. I would like to thank Prof. Dr. rer. nat. Werner E. G. Müller and Mrs. Renate Steffen, Institute of Physiological Chemistry and Pathobiochemistry, University of Mainz, for carrying out the cytotoxicity assays, Prof. Dr. rer. nat. M. U. Kassack and Dr. Alexandra Hamacher, Institut für Pharmazeutische und Medizinische Chemie, Düsseldorf for carrying out the cytotoxicity assays, Dr. Michael Kubbutat, ProQinase GmbH, Freiburg, Germany for conducting the protein kinase inhibition assays and Prof. Ute Hentschel Humeida, Julius-von-Sachs-Institut für Biowissenschaften, University of Würzburg, for carrying out the antibacterial and antifungal assays. I am grateful to Dr. Tibor Kurtán (Department of Organic Chemistry, University of Debrecen, Hungary) and his coworkers for their splendid CD calculation. My special thank to Mrs. Andrea Bieck for carrying out of my inscription at the University of Wuppertal, her continuous encouragement and her kind recommendation. Many thanks to technical assistants of : - the Institute of Pharmacological Biology and Biotechnology, Heinrich-Heine-Universität, Düsseldorf (Mrs. Miljanovic Simone, Mrs. Katja Friedrich, Mrs. Waltraud Schlag, Mr. Klaus Dieter Jansen, Mrs. Eva Müller, Mrs Goldbach-Gecke Heike) who never gave up providing assistance and continuously supplied me with the laboratory equipment and solvents during my laboratory work. - the Institute of Organic Chemistry, Bergische Universität, Wuppertal (Mrs. Simone Bettinger, Mr. Jürgen Dönecke, Mrs. Ilka Polanz, Mr. Andreas Siebert) for carrying out the 600 MHz NMR measurements, HR-MS experiments, and for their kind and generous help. iv Acknowledgement - the Laboratory of Pharmacokinetic, University Mohammed V, Rabat (Mrs. Hayat Houari and Mr. Ahmed Lafrouhi) for their sense of responsibility and their ongoing assistance. Many thanks to Mr. Andreas Marmann and Ms. Lena Hammerschmidt, who helped me much during my first adaptation months in the institute, introduced me to the variety of isolation techniques, and explained to me regularly the basic NMR spectra interpretation and structure elucidation, respectively. I would like to extend my thanks to my past and present colleagues and lab-mates Prof. Ag. Bouchra Meddah, Mrs. Mouna Elnekity, Ms. Nisrin Benayad, Ms. Mariam Mousa, Ms. Mounia Rochdi, Mr. Bouchra Faridi, Mr. Nabil Khouya, Ms. Violeta El Alami, Mr. Moustapha El Amrani, Mr. Weaam N. E. Ebrahim, Mr. Ilias Marmouzi, Mrs. Muharini Rini, Mr. Harvé Sergi Akoné and all the others for the nice multicultural time I spent with them, for their help and assistance whenever I needed it. My special thanks to my friends all over the world Mrs. Zineb Essakali, Mrs. Ruwayda Al Sardi, Mrs. Houda Damour, Mrs. Raha S. Orfali, Mrs. Meryem Carreau and Mrs. Jihane Laafi who encouraged me to stand and never surrender during my difficulties and always listen to me when I needed any moral support. Last but not least, my gratitude, thankfulness, and my gread indebtedness to my Husband Yassir Benmessaoud, my father Taibi Kabbaj, my mother Dikra Yamani, my sister and brother Khadija and Ismail Kabbaj, my stepfather Mohammed Benmessaoud, stepmother Hayat El Jirari, stepsisters Fatima Ezzahra Bemessaoud and Safiya Khalidi, stepbrothers Yassir Nadeif, Mehdi Benmessaoud and Rachid Maakoul and both the Kabbaj and Benmessaoud Families specially my aunt Atika Kabbaj, for their unfailing love, spiritual support and everlasting prayers. v Zusammenfassung Zusammenfassung Heilpflanzen und endophytische Pilze produzieren Naturstoffe mit