In Vitro Antiplasmodial Activity of Cochlospermum planchonii and its Phytomedicines
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1 In Vitro Antiplasmodial Activity of Cochlospermum planchonii and its Phytomedicines A. Lamien-Meda1, K. König1, M. Mischka1, R. Meda2, M. Compaoré3, M. Kiendrebeogo3, H.P. Fuehrer4, H. Nödl5, J. Novak1 1 Institute of Animal Nutrition and Functional Plant Compounds, University of Veterinary Medicine Vienna, Austria 2 UFR/ST, Université Polytechnique de Bobo-Dioulasso, Burkina Faso 3 Laboratory of Biochemistry and Chemistry (LABIOCA), UFR/SVT, University of Ouagadougou, Burkina Faso 4 Institute of Parasitology, University of Veterinary Medicine Vienna, Veterinaerplatz 1, Austria 5 Institute of Specific Prophylaxis and Tropical Medicine, Medical University of Vienna, Vienna, Austria
2 Research background MEAMP Project Focus on the endangered antimalarial medicinal plants from Burkina Faso: Malaria is leading cause of infantile death Malaria constitutes a public health preoccupation for the whole population Source: WHO Malaria Report 2011/2013
3 Recent Antimalarial Natural Products from Plants alkaloids 18 quassinoids 23 sesquiterpenes 27 triterpenoids 21 flavonoids/xanthones 9 quinones 25 miscellaneous compounds indole alkaloids (with high antiplasmodial activity) antiplasmodial natural products: Alkaloids, terpenes, quassinoids, flavonoids, limonoids, chalcones, peptides, xanthones, quinones, coumarins, miscellaneous compounds Saxena et al. (2003) Frederich et al. (2008) Kaur et al. (2009)
4 Cochlospermum planchonii Hook. f. ex Planch. (Bixaceae) Rhizome traditionally used for the treatment of fevers, jaundice and malaria in West African countries The phytomedicine N Dribala (rhizome of ): The active compounds of the plant still remain unknown
5 Cochlospermum planchonii Hook. f. ex Planch. (Bixaceae) Why? Survey results (APPEAR project-75)
6 Phytomedicines C.planchonii N dribala C.Planchonii root Saye
7 Extraction, Partition, Chromatography MeOH Ethanol Aceton CH2Cl2 Water
8 Continuous P. falciparum Culture Sterile hood (laminar flow); incubator, 37 C; gas mixture (5% O 2, 5% CO 2, 90% N 2 ); liquid nitrogen tank; Microscope with oil immersion objective Cell culture flasks RPMI 1640 Medium Erythrocyte concentrate, preferably 0- Human serum, preferably A+, AB+ or corresponding to RBCs Giemsa stain Parasite used for testing: chloroquine sensitive 3D7
9 HRP2 ELISA
10 Development of Quality Control System
11 HPLC (High Pressure Liquid Chromatography) Profile
12 HPLC (High Pressure Liquid Chromatography) Profile TLC:EtOAC/MeOH /H 2 O (77/13/10) Anisaldehyde/ visible Cochloxanthine Dihydrocochloxanthine Cochloxanthine Fractions of Cochlospermum plant material from MPLC (Aceton/Hexane 3/7; Acetic acid 0.1%) Dihydrocochloxanthine HPLC and structure elucidation by Nuclear Magnetic Resonance (NMR) spectroscopy
13 HPLC (High Pressure Liquid Chromatography) Profile TLC:EtOAC/MeOH /H 2 O (77/13/10) Anisaldehyde/ visible Cochloxanthine Dihydrocochloxanthine Cochloxanthine Fractions of Cochlospermum plant material from MPLC (Aceton/Hexane 3/7; Acetic acid 0.1%) Dihydrocochloxanthine HPLC and structure elucidation by Nuclear Magnetic Resonance (NMR) spectroscopy Are the two compounds active against malaria?
14 HPLC (High Pressure Liquid Chromatography) Profile TLC:EtOAC/MeO H/H 2 O (77/13/10) Anisaldehyd e/visible Cochloxanthine Dihydrocochloxanthine Cochloxanthine Fractions of Cochlospermum plant material from MPLC (Aceton/Hexane 3/7; Acetic acid 0.1%) rhizome Dihydrocochloxanthine HPLC and structure elucidation by Nuclear Magnetic Resonance (NMR) spectroscopy IC50 (µg/ml) Cochloxanthine 6.3 Dihydrocochloxanthine 6.6
15 MeOH: DH Cochloxanthine/Cochloxanthine Quality control
16 Ethanol: DH Cochloxanthine/Cochloxanthine Quality control
17 Quality control Acetone: DH Cochloxanthine/ Cochloxanthine
18 Quality control CH2Cl2: DH Cochloxanthine/Cochloxanthine
19 Quality control H2O: DH Cochloxanthine/Cochloxanthine
20 Why the decoction is active?
21 Cochlospermum: Antiplasmodial Activity IC50 (µg/ml) H2O H2O H2O MeOH EtOH CH2Cl2 Aceton Saye N'dribala CP2
22 Cochlospermum: Antiplasmodial Activity IC50 (µg/ml) H2O H2O H2O MeOH EtOH CH2Cl2 Aceton Saye N'dribala CP2 MeOH EtOH CH2Cl2 Aceton
23 Cochlospermum: Antiplasmodial Activity IC50 (µg/ml) H2O H2O H2O MeOH EtOH CH2Cl2 Aceton DCM EAF DH cochlox. cochlox. Saye N'dribala CP2 CP MeOH EtOH CH2Cl2 Aceton
24 Cochlospermum: Antiplasmodial Activity 60,0 50,0 40,0 30,0 20,0 10,0 0,0 26,9 39,2 56,8 43,3 IC50 (µg/ml) 50,9 50,3 20,8 0,8 10,9 6,6 6,4 H2O H2O H2O MeOH EtOH CH2Cl2 Aceton DCM EAF DH cochlox. cochlox. Saye N'dribala CP2 CP MeOH EtOH CH2Cl2 Aceton
25 Cochlospermum: Antiplasmodial Activity IC50, IC90, IC99 (µg/ml) 120 IC50 IC90 IC H2O H2O H2O MeOH EtOH CH2Cl2 Aceton DCM EAF DH cochlox. Saye N'dribala CP2 CP cochlox.
26 Marker Compounds for Quality Control, N dribala, Saye Cochloxanthine & Dihydrocochloxanthine UV spectra Cochloxanthine Dihydrocochloxanthine
27 Research Perspectives Further investigation on the antiplasmodial activity of : Fractions and isolated compounds from active fractions Investigation of the mode of action of the fractions on the parasite life stages. Cytotoxity of the the active fractions and/ or compounds
28 Acknowledgement: Austrian Partnership Programme in Higher Education and Research for Development APPEAR project-75 ( ) Prof. Harald Noedl Dr. Zephirin Dakuyo Aline Lamien Meda Johannes Novak Students: Martina Mischka Anna Karnthaler Johannes Klinger Karin König (BOKU) Associate Partner: Hans Peter Fuehrer Thomas Pacher Tamara Kolodziej Markus Bacher (BOKU) Associate Partners: University of Pretoria (Prof. K. ELOFF), University of Graz (Prof. Dr. R. BAUER), University of Lausanne (Dr. B. GRAZ), University of Oxford (Dr. M. WILLCOX), ULB Belgium (Prof. P. DUEZ) DI Elke Stinnig Programme Manager Appear Martin Kiendrebeogo Moussa Compaoré Roland Meda Student: Orokia Traoré Issa Karama Associate Partner: Odile G. Nacoulma Jeanne Millogo Patrice Zerbo Lassina Sanou Souleymane Sanou Marc Olivier
29 Vielen Dank für Ihre Aufmerksamkeit
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