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

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

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

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

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)

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

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

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., 2012. Int. J. Health Res. 5(2): 47-52   Symposium, Edo State Polytechnic, Usen

(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

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

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

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

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

References Chakraborthy GS, Badujarand RS and Pardeshi CR. (2009). Analgesic activity of chloroform extract of Caesalpinia pulcherrima. Journal of Pharmaceutical Research 2:1199-1200.   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):104-110. 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):2201-2206. 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, 6312-6319. 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):7887-7902.

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