cochlospermum planchonii and its phytomedicines

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

http://www.oead.at/

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 http://events.ucsf.edu/downloads/malaria/map.html

http://events.ucsf.edu/downloads/malaria/map.html

Recent Antimalarial Natural Products from Plants

1993-2003 127 alkaloids 18 quassinoids 23 sesquiterpenes 27 triterpenoids 21 flavonoids/xanthones 9 quinones 25 miscellaneous compounds

Saxena et al. (2003)

2008 31 indole alkaloids (with high antiplasmodial activity)

Frederich et al. (2008)

1998-2008 266 antiplasmodial natural products: Alkaloids, terpenes, quassinoids, flavonoids, limonoids, chalcones, peptides, xanthones, quinones, coumarins, miscellaneous compounds

Kaur et al. (2009)

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 C. planchonii):

•

• The active compounds of the plant still remain unknown

Why C. planchonii? Survey results (APPEAR project-75)

Cochlospermum planchonii Hook. f. ex Planch. (Bixaceae)

Phytomedicines

C.planchonii

N’dribala

Saye

C.Planchonii root

Extraction, Partition, Chromatography

MeOH Ethanol Aceton CH2Cl2 Water

Continuous P. falciparum Culture

• Sterile hood (laminar flow); incubator, 37°C; gas mixture (5% O2, 5% CO2, 90% N2); 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

HRP2 ELISA

Development of Quality Control System

C. planchonii HPLC (High Pressure Liquid Chromatography) Profile


Cochloxanthine

Dihydrocochloxanthine

TLC:EtOAC/MeOH/H2O (77/13/10)

Anisaldehyde/visible

Fractions of Cochlospermum plant material from MPLC (Aceton/Hexane 3/7; Acetic acid 0.1%)


Cochloxanthine

HPLC and structure elucidation by Nuclear Magnetic Resonance (NMR) spectroscopy


Cochloxanthine






Cochloxanthine


Are the two compounds active against malaria?

Cochloxanthine







Cochloxanthine


IC50 (µg/ml)

Cochloxanthine 6.3

Dihydrocochloxanthine 6.6 C. planchonii rhizome

0

2000000

4000000

6000000

8000000

10000000

12000000

14000000

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

MeOH: DH Cochloxanthine/Cochloxanthine

C. planchonii Quality control

C. planchonii

0

2000000

4000000

6000000

8000000

10000000

12000000

14000000

16000000

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

Ethanol: DH Cochloxanthine/Cochloxanthine

C. planchonii


0

2000000

4000000

6000000

8000000

10000000

12000000

14000000

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 19 20 21 23 24 25 26 27 28 29

Acetone: DH Cochloxanthine/ Cochloxanthine

C. planchonii


0

1000000

2000000

3000000

4000000

5000000

6000000

7000000

8000000

9000000

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

CH2Cl2: DH Cochloxanthine/Cochloxanthine

C. planchonii


0

200000

400000

600000

800000

1000000

1200000

1400000

1600000

1800000

1 2 3 4 5 6 7 8 9 12 13 14 15 18 24 25 28 29

H2O: DH Cochloxanthine/Cochloxanthine

C. planchonii


Why the decoction is active?

Cochlospermum: Antiplasmodial Activity C. planchonii

0 10 20 30 40 50 60

H2O H2O H2O MeOH EtOH CH2Cl2 Aceton

Saye N'dribala CP2

IC50 (µg/ml)


0 10 20 30 40 50 60

H2O H2O H2O MeOH EtOH CH2Cl2 Aceton

Saye N'dribala CP2

IC50 (µg/ml)

MeOH EtOH CH2Cl2 Aceton



27

39

57

43

51 50

21

1 11 7 6 0

10

20

30

40

50

60

H2O H2O H2O MeOH EtOH CH2Cl2 Aceton DCM EAF DH cochlox.

cochlox.

Saye N'dribala CP2 CP

IC50 (µg/ml)



26,9

39,2

56,8

43,3

50,9 50,3

20,8

0,8 10,9 6,6 6,4 0,0

10,0

20,0

30,0

40,0

50,0

60,0


cochlox.


IC50 (µg/ml)


0

20

40

60

80

100

120


cochlox.


IC50, IC90, IC99 (µg/ml)

IC50 IC90 IC99

C. planchonii, N‘dribala, Saye

Cochloxanthine & Dihydrocochloxanthine

Marker Compounds for Quality Control

UV spectra

Cochloxanthine


Research Perspectives

Further investigation on the antiplasmodial activity of C. planchonii: 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

Dr. Zephirin Dakuyo

Acknowledgement: Austrian Partnership Programme in Higher Education and

Research for Development – APPEAR project-75 (2012-2014)

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)

Prof. Harald Noedl

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

DI Elke Stinnig Programme Manager Appear

Vielen Dank für Ihre Aufmerksamkeit

cochlospermum planchonii and its phytomedicines

Documents