Malays. Appl. Biol. (2014) 43(1): 65–71
SYNERGISTIC EFFECTS OF GSK3 AND p38 MAPK INHIBITORS
ON GROWTH OF Plasmodium falciparum EX VIVO
MARHALIM, L., EMBI, N. and SIDEK, H.M.*
School of Biosciences and Biotechnology,
Faculty of Science and Technology,
Universiti Kebangsaan Malaysia, Bangi, 43600, Selangor
*Email:
This e-mail address is being protected from spambots. You need JavaScript enabled to view it
ABSTRACT
Pharmacological inhibitors of glycogen synthase kinase 3 (GSK3) and p38 mitogen-activated protein kinase (p38 MAPK), two kinases commonly associated with signaling within cells, have been shown to suppress in vivo or ex vivo growth of plasmodial sp. Here we evaluated antiplasmodial activities ex vivo of four inhibitors against GSK3 [lithium chloride (LiCl), kenpaullone, (2’Z, 3’E)-6-bromoindirubin-3’-oxime (BIO) and SB216763]; and two against p38 MAPK (RWJ67657 and SB202190) individually and in combination preparatory to understanding the role of protein kinases in plasmodial development. The order of decreasing growth-suppressing potencies of the GSK3 inhibitors tested against Plasmodium falciparum 3D7 cultured ex vivo in erythrocytes was BIO (IC50=3.13 ?M) > kenpaullone (IC50 = 18.3 ?M) > SB216763 (IC50= 27.12 ?M) > LiCl (IC50= 25 468 ?M). The p38 MAPK inhibitor, RWJ67657, displayed an IC50 of 7.52 ?M against 3D7. SB202190 was less effective at inhibiting 3D7 displaying an IC50 of 14.80 ?M. When tested in combination, marked synergism was observed for combination of BIO or SB216763 with RWJ67657. In conclusion, GSK3 and MAPK inhibitors showed potent antiplasmodial activities. Synergistic effects observed between BIO or SB216763 and RWJ67657 in inhibiting plasmodial growth may implicate interaction between MAPK and GSK3 pathways and warrant further investigation.
Key words: GSK3, MAPK, inhibitors, Plasmodium falciparum, malaria
REFERENCES
Berenbaum, M.C. 1978. A method for testing for synergy with any number of agents. The Journal of Infectious Disease, 137: 122-130.
Brumlik, M.J., Nkhoma, S., Kious, M.J., Thompson III., G.R., Patterson, T.F., Siekerka, J.J., Anderson, T.J.C. & Curiel, T.J. 2011. Human p38 mitogen-activated protein kinase inhibitor drugs Plasmodium falciparum replication. Experimental Parasitology. 128: 170-175.
Canfield, C.J., Pudney, M. & Gutteridge W.E. 1995. Interactions of atovaquone with other antimalarial drugs against Plasmodium falciparum in vitro. Experimental Parasitology, 80: 373-381.
Chen, Z., Gibson, T. B., Robinson, F., Silvestro, L., Pearson, G., Xu, B.-E., Wright, A., Vanderbilt, C. & Cobb, M. H. 2001. MAP kinases. Chemical Reviews, 101: 2449-2476.
Cohen, P. 2002. Protein kinases--the major drug targets of the twenty-first century? Nature Reviews Drug Discovery 1: 309-315.
Cohen, P. & Frame, S. 2001. The renaissance of GSK3. Nature Reviews Molecular Cell Biology. 2: 769-776.
Ding, Q., Xia, W., Liu, J.C., Yang, J.Y., Lee, D.F., Xia, J., Bartholomeusz, G., Li, Y., Pan, Y., Li, Z., Bargou, R.C., Qin, J., Lai, C.C., Tsai, F.J., Tsai, C.H. & Hung, M.C. 2005. Erk associates with and primes GSK-3? for its inactivation resulting in up-regulation of ?-catenin. Molecular Cell. 19: 159–170.
Doble, B.W. & Woodgett. J.R. 2003. GSK-3: tricks of the trade for a multitasking kinase. Journal of Cell Science, 116: 1175– 1186.
Dobson, S., Bracchi, V., Chakrabarti, D. & Barik, S. 2001. Characterization of a novel serine/threonine protein phosphatase (PfPPJ) from the malaria parasite, Plasmodium falciparum. Molecular and Biochemical Parasitolology, 115: 29-39.
Doerig, C. 2004. Protein kinases as targets for anti-parasitic chemotherapy. Biochimica et Biophysica Acta, 1697: 155–168.
Drocheau, 2004. Plasmodium falciparum Glycogen Synthase Kinase-3: Molecular Model, Expression, Intracellular Localisation and Selective Inhibitors, Biochimica et Biophysica Acta. 1697: 181-196.
Embi, N., Rylatt, D. B. & Cohen, P. 1980. Glycogen synthase kinase-3 from rabbit skeletal muscle; separation from cyclic-AMP-dependent protein kinase and phosphorylase kinase. European Journal of Biochemistry. 107: 519–527.
Hommes, D.W., Peppelenbosch, M.P. & Van Deventer, S.J.H. 2003. Mitogen activated protein (MAPK) kinase signal transduction pathways and novel anti-inflammatory targets. GUT. 52: 144-151
Gupta, S., Thapar, M.M., Wernsdorfer, W.H. & Bjorkman, A. 2002. In vitro interactions of artemisinin with atovaquone, quinine, and mefloquine against Plasmodium falciparum. Antimicrobial Agents and Chemotherapy, 46: 1510-1515.
Kebeneil, J.S., Ndalut, P.K. & Sabah, A.O. 2004. Synergism of artemisinin with abyssinone –V from Erythrina abyssinica (Lam. ex) against Plasmodium falciparum parasites: A potential anti-malarial combination therapy. Journal of Medicinal Plants Research. 5 :1355-1360.
Klein, P.A & Melton, D.A.1996. A molecular mechanism for the effect of lithium on development. Proceedings of the National Academy of Sciences, 93: 8455-8459.
Kim, E.K. & Choi, E.J. 2010. Pathological roles of MAPK signaling pathways in human disease. Biochimica et Biophysica Acta. 1802: 396-405
Kirshennboim, N., Plotkin, B., Ben, Shlomo, S., Kaidanovich-Beilin, O. & Eldar-Finkelman, H. 2004. Lithium-mediated phosphorylation of glycogen synthase kinase-3 involves PI3Kinase-dependent activation of protein kinaseC- alpha. Journal of Molecular Neuroscience. 24: 199–207.
Lawrence, M.C., Jivan, A., Shao, C., Duan, L., Goad, D., Zaganjor, E., Osborne, J., McGlynn, K., Stippec, S., Earnest, S., Chen, W. & Cobb, M.H. 2008. Review: The roles of MAPKs in disease. Cell Research. 18: 436-442.
Makler, M.T., Ries, J.M., Williams, J.A., Bancroft, J.E., Piper, R.C., Gibbons, B.L. & Hinrichs, D.J. 1993. Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. The American Journal of Tropical Medicine and Hygiene, 48: 739–741.
Meijer, L., Skaltsounis, A.L., Magiatis, P., Polychronopoulos, P., Knockaert, M., Leost, M., Ryan, X.P., Vonica, C.A., Brivanlou, A., Dajani, R., Crovace, C., Tarricone, C., Musacchio, A., Roe, S.M., Pearl, L. & Greengard, P. 2003. GSK-3-Selective Inhibitors Derived from Tyrian Purple Indirubins. Cell press. 10: 1255-1266
Meijer, L., Flajolat, M. & Greengard, P. 2004. Pharmacological inhibitors of glycogen synthase kinase 3. Trends in Pharmacological Sciences. 25 (9) : 471-480.
Nurul Aiezzah, Z., Noor, E. & Hasidah, M. S. 2010. Suppression of Plasmodium berghei parasitemia by LiCl in an animal infection model. Tropical Biomedicine. 27 (3): 624-631.
Ojo, K.K., Gillespie, J.R., Riechers, A.J., Napuli, A.J., & Verlinde, C.L. 2008. Glycogen synthase kinase 3 is a potential drug target for African trypanosomiasis therapy. Antimicrobial Agents Chemotherapy. 52: 3710–3717.
Pfister, D.D., Burkard, G., Morand,S., Renggli, C.K., Roditi, I. & Vassella, E. 2006. A Mitogen-Activated Protein Kinase Controls Differentiation of Bloodstream Forms of Trypanosoma brucei. Eukaryotic Cell, 5: 1126–1135
Radfar, A., Mendez, D., Moneriz C, Linares, M., Marin-Garcia P, Puyet, A., Diez, A. & Bautista, J.M. 2009. Synchronous culture of Plasmodium falciparum at high parasitemia levels. Nature Protocols, 4: 1899-1915.
Schutz, S.V., Schrader, A.J., Zengerling, F., Genze, F., Cronauer M.V. & Schrader, M. 2011. Inhibition of Glycogen Synthase Kinase-3? Counteracts Ligand-Independent Activity of the Androgen Receptor in Castration Resistant Prostate Cancer. PLOS ONE, 6: e25341
Skinner, T.S., Manning, L.S, Johnston, W.A. & Davis, T.M.E. 1996. In vitro Stage-specific Sensitivity of Plasmodium falciparum to Quinine and Artemisinin Drugs. International Journal for Parasitology, 26: 519-525
Thornton, T.M., Pedraza-Alva, G., Deng, B., Wood, C.D., Aronshtam, A., Clements, J.L., Sabio, G., Davis, R.J., Matthews, D.E., Doble, B. & Rincon, M. 2008. Phosphorylation by p38 MAPK as an alternative pathway for GSK3beta inactivation. Science. 320 (5876): 667-670.
Winstanley. P.A. 2000. Chemotherapy for P. falciparum malaria: The Armoury, The Problems And The Prospects. Parasitology Today, 16: 146–153.
Wei, S., Marchesb, F., Daniela, B., Sondac, S., Heidenreichd, K. & Curiela, T. 2002. Pyridinylimidazole p38 mitogen-activated protein kinase inhibitors block intracellular Toxoplasma gondii replication. International Journal for Parasitology. 32: 969–977.
World Health Organization (WHO). 2012. World malaria report 2012. 195 pp.
Xingi E., Smirlis D. & Myrianthopoulos, V. 2009. 6-Br-5-methylindirubin-3 ' oxime (5-Me-6-BIO) targeting the leishmanial glycogen synthase kinase-3 (GSK-3) short form affects cell-cycle progression and induces apoptosis-like death: Exploitation of GSK-3 for treating leishmaniasis. International Journal for Parasitology, 39:1289–303.
