Heme Polymerization Inhibition by Tithonia diversifolia (Hemsley) A.Gray Leaves Fractions as Antiplasmodial Agent and Its Cytotoxicity on Vero Cells

https://doi.org/10.22146/mot.36476

Rul Afiyah Syarif(1*), Mustofa Mustofa(2), Ngatidjan Ngatidjan(3), Mae Sri Hartati Wahyuningsih(4)

(1) Department of Pharmacology and Therapy, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada
(2) Department of Pharmacology and Therapy, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada
(3) Department of Pharmacology and Therapy, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada
(4) Department of Pharmacology and Therapy, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada
(*) Corresponding Author

Abstract


Previous research revealed that the extracts and fractions of Tithonia diversifolia (Hemsley) A.Gray leaves had antiplasmodial activity in vitro. For further development as an antiplasmodial agent, the mechanisms of action and safety of compounds are important to disclose. Heme polymerization inhibition is one of the main targets of antiplasmodial action. The aim of the study was to investigate the activity of T. diversifolia fractions in inhibiting heme polymerization and its cytotoxic effect on Vero cells. Heme polymerization inhibition assay from Bassilico and cytotoxic test on Vero cell using MTT method were conducted for three fractions (F5, F6, and F7) of T. diversifolia leaves. The inhibitory activity of heme polymerization expressed as IC50 and cytotoxicity effect expressed as CC50 were determined by probit analysis. The best heme polymerization inhibition activity was F5 with IC50 = 162.20 ± 57.81 μg/mL followed by F6 and F7 with IC50 216.30 ± 26.56 and 231.54 ± 44.26 μg/mL respectively. All the fractions had a low cytotoxic effect with CC50 for F5, F6, and F7 were over than 100, 34.81 ± 9.94 and 56.26 ± 6.73 μg/mL, respectively and the toxicity index fraction is below 10 or categorized as low selectivity. Conclusion: The fraction of T. diversifolia inhibited heme polymerization in vitro and had low cytotoxic effect on Vero cells but no selective toxicity. Further research using pure compounds may improve its selectivity.


Keywords


Tithonia diversifolia; antiplasmodial; heme polymerization; cytotoxicity

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References

Bassilico, N., Pagani, E., Monti, D., Olliaro, P., and Taramelli, D., 1998, A microtitre-based method for measuring the Haem Polimerization Inhibitory Activity (HPIA) of antimalarial drugs, J. Antimicrob. Chemother. 42:55-60.

Becker, K., Tilley, L., Vennerstrom, J.L., Roberts, D., Rogerson, S., Ginsburg, H. 2004. Oxidative stress in malaria parasite-infected erythrocytes: host-parasite interactions. Int. J. Parasitol. 34(2):163-89.

Campanale, N., Nickel, C., Daubenberger, C.A., Wehlan, D.A., Gorman, J.J., Klonis, N., Becker, K., Tilley, L. 2003. Identification and characterization of heme-interacting proteins in the malaria parasite, Plasmodium falciparum. J. Biol. Chem. 278(30): 27354-27361.

Carney, C.K., Harry, S.R., Sewell, S.L.,.Wright, D.W. 2007. Detoxification biominerals. Top. Curr. Chem. 270: 155–185.

Chong, C.R and Sullivan, D.J. 2003. Inhibition of heme crystal growth by antimalarials and other compounds: implications for drug discovery. Biochem. Pharmacol. 66: 2201–2212.

Coronado, L.M., Nadovich, C.T., and Spadafora, C. 2014. Malarial hemozoin: from target to tool. Biochim. Biophys. Acta. 1840(6): 2032–2041.

Egan, T.J., Hunter, R., Kaschula, C.H., Marques, H.M., Misplon, A., Walden, J. 2000. Structure-function relationships in aminoquinolines: effect of amino and chloro groups on quinoline-hematin complex formation, inhibition of beta-hematin formation, and antiplasmodial activity. J. Med. Chem. 43(2):283-291.

Ehlgen, F., Pham, J.S., Koning-Ward, T., Cowman, A.F., Ralp, S.A. 2012. Investigation of the Plasmodium falciparum food vacuole through inducible expression of the Chloroquine Resistance Transporter (PfCRT). PLoS ONE. 7(6): 1-12.

Ghantous, A., Gali-Muhtasib, H., Vuorela, H., Saliba, N.A., and Darwiche, N. 2010. What made sesquiterpene lactones reach cancer clinical trials? Drug Discovery Today. 15 (15/16): 668-678.

Goffin, E., Ziemons, E., de Mol, P., de Madureira Mdo, C., Martins, A.P., da Cunha, A.P., Phillippe, G., Tits, M., Angenot, L., and Frederich, M. 2002. In vitro antiplasmodial activity of Tithonia diversifolia and identification of its main active constituent : taginin C. Planta Med., 68(6): 543-5.

Huy, N.T., Mizunuma, K., Kaur, K., Nhien, N.T.T., Jain, M., Uyen, D.T., Harada, S., Jain, R., and Kamei, K. 2007. 2-tert-Butyl-8-quinolinamines exhibit potent blood schizontocidal antimalarial activity via inhibition of heme crystallization. Antimicrob. Agents Chemother. 51(8): 2842-2847.

Jenett-Siems, K., Mockenhaupt, F.P., Bienzle, U., Gupta, M.P., and Eich, E. 1999. In vitro antiplasmodial activity of Central American medicinal plants. Trop. Med. Intl. Health., 4(9): 611-615.

Kumar, S., Guha, M., Choubey, V., Maity, P., Bandyopadhyay, U. 2007. Minireview antimalarial drugs inhibiting hemozoin (β-hematin) formation: A mechanistic update. Life Sci. 80: 813–828.

Kurosawa,Y., Dorn, A., Kitsuji-Shirane, M., Shimada,H., Satoh,T., Matile, H., Hofheinz, W., Masciadri, R., Kansy, M., and Ridley, R.G. 2000. Hematin polymerization assay as a high-throughput screen for identification of new antimalarial pharmacophores. Antimicrob. Agents Chemother. 44 (10): 2638–2644.

Lombardino, J.G. and Lowe, J.A. 3rd. 2004.The role of the medicinal chemist in drug discovery then and now. Nat. Rev. Drug Discov. 3(10): 853–862.

Nagababu, E., Chrest, F.J., Rifkind, J.M. 2003. Hydrogen-peroxide-induced heme degradation in red blood cells: the protective roles of catalase and glutathione peroxidase. Biochim. Biophys. Acta. 1620(1-3): 211-217.

Pandey, A.V. dan Chauhan, V.S. 1998. Heme polymerization by malarial parasites: a potential target for antimalarial drug development. Current Science. 75 (9): 911-918.

Syarif, R.A., Wahyuningsih, M.S.H., Mustofa, Ngatidjan. 2014. Aktivitas penghambatan Ekstrak terpurifikasi Daun Kembang Bulan (Tithonia diversifolia [Hemsley] A.Gray) pada Pertumbuhan Plasmodium falciparum dan Polimerase Heme. J. Indon. Med. Assoc. 64(5): 228-233.

Syarif, R.A., Wahyuningsih, M.S.H, Mustofa, Ngatidjan. 2018. Antiplasmodial activity and onset speed of growth inhibition of Tithonia diversifolia (Hemsley) A.Gray leave fractions on Plasmodium falciparum. Trop. J. Pharm. Res. under review.

Tada, H., Shiho, O., Kuroshima, K., Koyama, M., and Tsukamoto, K. 1986. An improved colorimetric assay for interleukin 2. J. Immunol. Methods. 93(2):157-165.

WHO. 2015. Guideline for the Treatment of Malaria. 3th Ed. 20 Avenua Appia, 1211 Geneva 27, Switzerland. WHO Press.





DOI: https://doi.org/10.22146/mot.36476

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