Antileishmanial Potential of Piper nigrum Seed Extracts against Leishmania donovani

Garima Chouhan¹, Mohammad Islamuddin¹, Farnaz Ahmad¹, Dinkar Sahal², Farhat Afrin^1,3*

¹Parasite Immunology Laboratory, Department of Biotechnology, Faculty of Science, Jamia Hamdard (Hamdard University), New Delhi, India.
²Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India.
³Department of Medical Laboratories Technology, Faculty of Applied Medical Sciences, Taibah University, Medina, KSA.

DOI: 10.4236/ojmm.2014.44025                

Abstract

Visceral leishmaniasis (VL) is a pestilent form of leishmaniasis that chiefly impinges the poorest sections of the society. The prototypical therapeutic interventions in vogue are handicapped due to toxicity and alarming increase in drug resistance. In the absence of vaccines, progressive emergence of HIV-VL co-infection and relapse in the form of post kala-azar dermal leishmaniasis, have fuelled the quest for alternative therapies. Herein, we report antileishmanial activity of Piper nigrum, which is endowed with multifarous medicinal properties. Hexane (PNH) and ethanolic (PNE) extracts of P. nigrum substantially inhibited the growth of Leishmania donovani promastigotes with 50% inhibitory concentration (IC₅₀) of 31.6 and 37.8 μg·ml^-1, respectively. Growth reversibility analysis revealed the leishmanicidal effect of PNH which caused cell shrinkage and flagellar disruption. In contrast, PNE treated promastigotes showed partial effect. PNH and PNE also abrogated the growth of intra-macrophagic Leishmania amastigotes with IC₅₀ of 14.6 and 18.3 μg·ml^-1, respectively. Anti-amastigote efficacy of PNH was accompanied by higher selectivity over host macrophages than PNE. Gas-Chromatography-Mass Spectrometry showed the presence of several secondary metabolites such as trans-β-caryophyllene, piperine, β-bisabolene and other sesquiterpenes in PNH and piperine, δ-(sup 9)-cis oleic acid and piperyline in PNE. Conclusively, our work revealed discernible antileishmanial activity of P. nigrum extracts.

Keywords

Visceral Leishmaniasis, Leishmania donovani, Piper nigrum, Antileishmanial, Plant Extracts

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Chouhan, G., Islamuddin, M., Sahal, D. and Afrin, F. (2014) Exploring the Role of Medicinal Plant Based Immunomodulators for Effective Therapy of Leishmaniasis. Frontiers in Immunology, 5, 193. http://dx.doi.org/10.3389/fimmu.2014.00193
[2]	Rahmatullah, M., Hasan, A., Parvin, W., Moniruzzaman, M., Khatun, A., Khatun, Z., Jahan, F.I. and Jahan, R. (2012) Medicinal Plants and Formulations Used by the Soren Clan of the SantalTribe in Rajsahi District, Bangladesh for Treatment of Various Ailments. African Journal of Traditional, Complementary and Alternative Medicine, 9, 350-359. http://dx.doi.org/10.4314/ajtcam.v9i3.8
[3]	Hermoso, A., Jiménez, I.A., Mamani, Z.A., Bazzocchi, I.L., Pinero, J.E., Ravelo, A.G. and Valladares, B. (2003) Antileishmanial Activities of Dihydrochalcones from Piper elongatum and Synthetic Related Compounds. Structural Requirements for Activity. Biorganic and Medicinal Chemistry, 11, 3975-3980. http://dx.doi.org/10.1016/S0968-0896(03)00406-1
[4]	Carrara, V.D.S., Cunha-Júnior, E.F., Torres-Santos, E.C., Corrêa, A.G., Monteiro, J.L., Demarchi, I.G., Lonardoni, M.V.C. and Cortez, D.A.G. (2013) Antileishmanial Activity of Amides from Piper amalago and Synthetic Analogs. Revista Brasiliera de Farmacognosia, 23, 447-454. http://dx.doi.org/10.1590/S0102-695X2013005000022
[5]	Sahi, S., Tewatia, P. and Ghosal, S. (2012) Leishmania donovani Pteridine Reductase 1: Comparative Protein Modeling and Protein-Ligand Interaction Studies of the Leishmanicidal Constituents Isolated from the Fruits of Piper longum. Journal of Molecular Modeling, 18, 5065-5073. http://dx.doi.org/10.1007/s00894-012-1508-y
[6]	Sen, R. and Chaterjee, M. (2011) Plant Derived Therapeutics for the Treatment of Leishmaniasis. Phytomedicine, 18, 1056-1069. http://dx.doi.org/10.1016/j.phymed.2011.03.004
[7]	Afrin, F., Rajesh, R., Anam, K., Gopinath, M., Pal, S. and Ali, N. (2002) Characterization of Leishmania donovani Antigens Encapsulated in Liposomes That Induce Protective Immunity in BALB/c Mice. Infection Immunity, 70, 6697-6706. http://dx.doi.org/10.1128/IAI.70.12.6697-6706.2002
[8]	Islamuddin, M., Farooque, A., Dwarakanath, B.S., Sahal, D. and Afrin, F. (2012) Extracts of Artemisia annua Leaves and Seeds Mediate Programmed Cell Death in Leishmania donovani. Jounal of Medical Microbiology, 61, 1709-1718. http://dx.doi.org/10.1099/jmm.0.049387-0
[9]	Islamuddin, M., Sahal, D. and Afrin, F. (2014) Apoptosis-Like Death in Leishmania donovani Promastigotes Induced by Eugenol-Rich Oil of Syzygium aromaticum. Journal of Medical Microbiology, 63, 74-85. http://dx.doi.org/10.1099/jmm.0.064709-0
[10]	Githinji, E.K., Irungu, L.W., Tonui, W.K., Rukunga, G.M., Mutai, C., Muthaura, C.N., Lugalia, R., Gikandi, G., Wainaina, C.W., Ingonga, J.M. and Wanjoya, A. (2010) In Vitro Effects of Warburgia ugandensis, Psiadia punctulata and Chasmanthera Dependes on Leishmania Major Promastigotes. African Journal of Traditional, Complementary and Alternative Medicine, 7, 264-275. http://dx.doi.org/10.4314/ajtcam.v7i3.54791
[11]	Benítez, J., Cavalcanti de Queiroz, A., Correia, I., Alves, M.A., Alexandre-Moreira, M.S., Barreiro, E.J., Lima, L.M., Varela, J., González, M., Cerecetto, H., Moreno, V., Pessoa, J.C. and Gambino, D. (2013) Newoxivanadium (IV) N-Acylhydrazone Complexes: Promising Antileishmanial and Antitrypanosomal Agents. European Journal of Medicinal Chemistry, 62, 20-27. http://dx.doi.org/10.1016/j.ejmech.2012.12.036
[12]	Soares, D.C., Portella, N.A., De S. Ramos, M.F., Siani, A.C. and Saraiva, E.M. (2013) Trans-β-Caryophyllene: An Effective Antileishmanial Compound Found in Commercial Copaiba Oil (Copaifera spp.). Evidence-Based Complementary and Alternative Medicine, 2013, Article ID: 761323. http://dx.doi.org/10.1155/2013/761323
[13]	Ferreira, C., Soares, D.C., Barreto-Junior, C.B., Nascimento, M.T., Freire-de-Lima, L., Delorenzi, J.C., Lima, M.E., Atella, G.C., Folly, E., Carvalho, T.M., Saraiva, E.M. and Pinto-da-Silva, L.H. (2011) Leishmanicidal Effects of Piperine, Its Derivatives, and Analogues on Leishmania amazonensis. Phytochemistry, 72, 2155-2164. http://dx.doi.org/10.1016/j.phytochem.2011.08.006
[14]	Osorio, E., Arango, G.J., Jiménez, N., Alzate, F., Ruiz, G., Gutiérrez, D., Paco, M.A., Giménez, A. and Robledo, S. (2007) Antiprotozoal and Cytotoxic Activities in Vitro of Colombian Annonaceae. Journal of Ethnopharmacology, 111, 630-635. http://dx.doi.org/10.1016/j.jep.2007.01.015
[15]	Odonne, G., Herbette, G., Eparvier, V., Bourdy, G., Rojas, R., Sauvain, M. and Stien, D. (2011) Antileishmanial Sesquiterpene Lactones from Pseudelephantopus spicatus, a Traditional Remedy from the Chayahuita Amerindians (Peru). Part III. Journal of Ethnopaharmacology, 137, 875-879.
[16]	Izumi, E., Ueda-Nakamura, T., Veiga Jr., V.F., Pinto, A.C. and Nakamura, C.V. (2012) Terpenes from Copaifera Demonstrated in Vitro Antiparasitic and Synergic Activity. Journal of Medicinal Chemistry, 55, 2994-3001. http://dx.doi.org/10.1021/jm201451h
[17]	Amiel, E., Ofir, R., Dudai, N., Soloway, E., Rabinsky, T. and Rachmilevitch, S. (2012) β-Caryophyllene, a Compound Isolated from the Biblical Balm of Gilead (Commiphora gileadensis), Is a Selective Apoptosis Inducer for Tumor Cell Lines. Evidence-Based Complementary and Alternative Medicine, 2012, 872394. http://dx.doi.org/10.1155/2012/872394
[18]	Kapil, A. (1993) Piperine: A Potent Inhibitor of Leishmania donovani Promastigotes in Vitro. Planta Medica, 59, 474. http://dx.doi.org/10.1055/s-2006-959737
[19]	Singh, I.P., Jain, S.K., Kaur, A., Singh, S., Kumar, R., Garg, P., Sharma, S.S. and Arora, S.K. (2010) Synthesis and Antileishmanial Activity of Piperoyl-Amino Acid Conjugates. European Journal of Medicinal Chemistry, 45, 3439- 3445. http://dx.doi.org/10.1016/j.ejmech.2010.04.033