1. Symposium, Edo State Polytechnic, Usen
Cassane Diterpenoids and Derivatives from the Roots of Caesalpinia pulcherrima with Selective Cytotoxic Activity Against Multiple Myeloma Cells
Osayemwenre Erharuyi, Rehan Imad, Scott Simanski, Paige Dickson, Thomas Kodadek, Abiodun Falodun1, and M. Iqbal Choudhary
Presented By  Osayemwenre Erharuyi
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Benin, Benin City, Nigeria.
Symposium, Edo State Polytechnic, Usen

2. INTRODUCTION
Nature as an attractive source of drugs, drug leads and new chemical entities (NCEs) can be found in millions of species of plants, animals, marine organisms and microbes. Natural products of plant origin have been used as foods and as medicines since human existence.
Symposium, Edo State Polytechnic, Usen

3. Caesalpinia pulcherrima
Caesalpinia pulcherrima in its natural habitat
Caesalpinia pulcherrima L. (family; Caesalpiniaceae) commonly called Pride of Barbados is an evergreen perennial shrub or small tree widely distributed in the tropics.
The plant has been used in ethnomedicine for a variety of ailments especially as a tonic, stimulant and abortifacient.
The plant is a rich source of diterpenoids of the cassane-type
(Zanin et al., 2012; Chakraborty et al., 2009)
Symposium, Edo State Polytechnic, Usen

4. Aim of Study
To further evaluate the biological significance of this plant, we investigated the cancer cell cytotoxic effects of the cassane diterpenoids previously isolated from the root bark of the plant.
Symposium, Edo State Polytechnic, Usen

5. The powdered plant materials were extracted by maceration in methanol.
Materials and Methods
The powdered plant materials were extracted by maceration in methanol.
chromatography
Cassane Diterpenoids
Crude extract
(CPR)
Chloroform fraction (CP-Ch)
Petroleum ether fraction (CP-Pe)
Ethyl acetate fraction (CP-Et)

6. (Erharuyi et al., 2016)
Symposium, Edo State Polytechnic, Usen

7. (Erharuyi et al., 2017)
Symposium, Edo State Polytechnic, Usen

8. Cytotoxic screening
The diterpenoids were evaluated for their cytotoxic effects on three cancer cell lines;
cervical cancer (HeLa),
lung cancer (A549),
multiple myeloma (MM.1S) and
the normal human embryonic kidney (HEK293T) cells
The assay was carried out using the CellTitre-Glo assay as previously described (Trader et al., 2015).
***P < 0.001
Figure 1: Cell cycle measurement of MCF-7 cells in treated (JMR) and untreated (DMSO-Control) conditions
Falodun et al., Int. J. Health Res. 5(2): 47-52
Symposium, Edo State Polytechnic, Usen

9. (Percentage inhibition)
Results and Discussion
Table 1: Cytotoxic activity (Percentage inhibition) of compounds 1-11.
Compound
Cytotoxicity
(Percentage inhibition)
HeLa
A543
MM.1S
HEK239T 1
41.98 ± 3.96
65.45 ± 1.44
80.32
60.56 ± 1.76 2
86.24 ± 2.45
63.24± 1.67
96.7± 1.23
89.46± 3.68 3
45.65 ± 0.21
77.02 ± 0.31
75.64 ± 1.30
69.08 ± 1.52 4
22.43 ± 4.70
88.42 ± 0.40
82.24 ± 0.60
78.96± 3.43 5
50.34 ± 1.44
40.24 ± 1.44
48.79± 2.44
40.23± 1.56 6
90.98± 1.24
89.23 ± 1.40
98.79± 2.44
90.23± 1.56 7
50.67± 1.59
14.50 ± 0.79
18.50 ± 1.90
10.03 ± 1.56 8
19.13 ± 0.88
18.12 ± 1.53
20.34 ± 1.98
15.3 ± 1.1 9
96.49 ± 1.39
90.09 ± 1.12
99.59 ± 0.26
94.7 ± 4.8 10
31.74 ± 1.68
27.61 ± 2.49
67.60 ± 2.0
58.90 ± 2.56 11
50.78
48.70 ± 1.06
50.78 ± 3.54
52.77 ± 2.55
Bortexomib
97.89 ± 0.10
90.22 ± 0.04
99.30 ± 0.02
89.22 ± 1.66 *
Symposium, Edo State Polytechnic, Usen

10. Results and Discussion
Table 2: Cytotoxic activity of compounds 2, 6 and 9 against the selected cell lines.
Compound
IC50 (µM)
HeLa
A569
MM.1S
HEK293T 2
21.43
20.17
19.64
31.28 6
28.98
55.12
7.44
24.61 9
20.01
22.35
1.07
7.55
Symposium, Edo State Polytechnic, Usen

11. Results and Discussion
Figure 1: (A) Selective toxicity of a natural product-derived cassane diterpenoid (9) to multiple myeloma cells. (B) Synergistic cytotoxic activity of compound 9 with bortezomib.
Symposium, Edo State Polytechnic, Usen

12. Conclusion
The present study has shown a potential of the cassane diterpenoids and its derivatives as a cytotoxic agent against cancer cells particularly against multiple myeloma either as a mono-therapy or in combination with known cytotoxic drugs.
Symposium, Edo State Polytechnic, Usen

13. THE FULBRIGHT FOREIGN STUDENT PROGRAM THE TWAS-ICCBS FELLOWSHIP
acknowledments
Department of Pharmaceutical Chemistry,
University of Benin, Nigeria
Prof. A. Falodun
Department of Chemistry and Cancer Biology,
The Scripps Research Institute, Jupiter, Florida, U.S.A.
Prof. Thomas Kodadek
Dr. Patrick McEnaney
Dr. Scott Simanski
Dr. Paige Dickson
International Centre for Chemical and Biological Sciences, University of Karachi, Pakistan.
Prof. M. Iqbal Choudhary
THE FULBRIGHT FOREIGN STUDENT PROGRAM
THE TWAS-ICCBS FELLOWSHIP
Symposium, Edo State Polytechnic, Usen

14. References
Chakraborthy GS, Badujarand RS and Pardeshi CR. (2009). Analgesic activity of chloroform extract of Caesalpinia pulcherrima. Journal of Pharmaceutical Research 2:
Erharuyi O, Adhikari A, Falodun A, Jabeen A, Imad R, Ammad M, Choudhary MI, GÖren N. (2017). Cytotoxic, anti-inflammatory, and leishmanicidal activities of diterpenes isolated from the roots of Caesalpinia pulcherrima. Planta Medica 83(01/02):
Erharuyi O, Adhikari A, Falodun A, Imad R and Choudhary MI. (2016). Derivatization of cassane diterpenoids from Caesalpinia pulcherrima (L.) Sw. and evaluation of their cytotoxic and leishmanicidal activities. Tetrahedron Letters 57(20):
Trader DJ, Simanski S, Kodadek T. (2015). A Reversible and Highly Selective Inhibitor of the Proteasomal Ubiquitin Receptor Rpn13 Is Toxic to Multiple Myeloma Cells. J. Am. Chem. Soc. 137,
Zanin JL, de Carvalho BA, Martineli PS, dos Santos MH, Lago JH, Sartorelli P, Viegas C Jr, Soares MG.
The genus Caesalpinia L. (Caesalpiniaceae): phytochemical and pharmacological characteristics. Molecules 2012;17(7):

15. THANK YOU
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