Azerigyik, Faustus AkankperiwenAmoa-Bosompem, MichaelTetteh, ThelmaAyertey, FrederickAntwi, Ama NyamekyeOwusu, Kofi Baffour-AwuahDadzie, Kofi KwofieDjameh, Georgina IsabellaTetteh-Tsifoanya, MarkIwanaga, ShiroAppiah, Alfred AmpomahOhta, TomoeUto, TakuhiroShoyama, YukihiroOhta, NobuoGwira, Theresa ManfulOhashi, Mitsuko2020-01-022020-01-022018-11Azerigyik Faustus Akankperiwen, Amoa-Bosompem Michael, Tetteh Thelma, Ayertey Frederick, Antwi Ama Nyamekye, Owusu Kofi Baffour-Awuah, Dadzie Kofi Kwofie, Djameh Georgina Isabella, Tetteh-Tsifoanya Mark, Iwanaga Shiro, Appiah Alfred Ampomah, Ohta Tomoe, Uto Takuhiro, Shoyama Yukihiro, Ohta Nobuo, Gwira Theresa Manful, Ohashi Mitsuko. In vitro Mechanistic Assays of Tetracyclic Iridoid Compounds Isolated from Morinda lucida Benth in Leishmania species. European Journal of Medicinal Plants. 2018 Nov; 25(4): 1-142231-0894http://imsear.searo.who.int/handle/123456789/189430Aims: This study investigates the activity of tetracyclic iridoid compounds against Leishmania spp. and the mechanism(s) of action. Study Design: An experimental study. Place and Duration: Department of Parasitology, Noguchi Memorial Institute for Medical Research, between September 2017 and July 2018. Methodology: The 50 % inhibitory concentration (IC50) of compounds against Leishmania donovani and L. major promastigotes were determined after 48 hours of incubation using the Alamar blue. Cytotoxicity of compounds was determined against cell lines using MTT assay. The anti-amastigote activity of compounds was further assessed by DAPI (4′,6-diamidino-2-phenylindole) staining. The mechanism of cell death induced by compounds was determined using nexin assay. Mitosis, cytokinesis and morphometry were monitored by DAPI and Kinetoplastid Membrane Protein (KMP) staining. Cell cycle arrest induced by compounds was analyzed by FACS. Results: Molucidin and ML-F52 inhibited the growth of promastigote in L. donovani (Molucidin; IC50 = 2.94±0.60 µM, ML-F52; IC50 = 0.91±0.50 µM) and L. major (Molucidin; IC50 = 1.85± 0.20 µM, ML-F52; IC50 = 1.77± 0.20 µM). ML-F52 had a 10-fold cytotoxic effect on parasites relative to normal cell lines. Against intracellular forms, Molucidin and ML-F52 inhibited intracellular amastigote replication and infectivity. Amphotericin B, Molucidin and ML-F52, induced a dose-dependent apoptotic effect on promastigotes. Although no change in KMP-11 expression was observed, iridoids inhibited cell division and morphological changes in promastigote cultures. Molucidin and ML-F52 induced apoptotic mechanism of cell death, inhibited cytokinesis and induced phenotypic changes in promastigotes. Molucidin further induced ‘’nectomonad-like’’ forms and loss of kDNA, ML-F52 induced ‘cell-rounding’ with loss of flagellum. Molucidin also induced cell growth arrest at G2-M phase (54.5 %). A significant induction of apoptosis (P = .05) was shown by an enhanced peak in the sub-G1 confirming the apoptotic inducing properties of iridoids. Conclusion: This study shows the anti-leishmania activity of tetracyclic iridoids which could be further investigated for the development of new chemotherapy against Leishmaniasis.Leishmania donovaniLeishmania majorIn vitro screeningmedicinal plantsTetracyclic iridoidsMorinda lucidaApoptosisJournal ArticleIndiaWest African Centre for Cell Biology and Infectious Pathogens (WACCBIP), Department of Biochemistry, Cell and Molecular Biology (DBCMB), University of Ghana, Legon, Ghana and Noguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, GhanaNoguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, Ghana and Section of Environmental Parasitology, Faculty of Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanNoguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, GhanaCentre for Plant Medicine Research, P.O.Box 73, Mampong - Akuapem, GhanaNoguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, GhanaNoguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, GhanaSection of Environmental Parasitology, Faculty of Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanNoguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, GhanaNoguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, GhanaSection of Environmental Parasitology, Faculty of Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanCentre for Plant Medicine Research, P.O.Box 73, Mampong - Akuapem, GhanaFaculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch, Sasebo, Nagasaki 859-3298, JapanFaculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch, Sasebo, Nagasaki 859-3298, JapanFaculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch, Sasebo, Nagasaki 859-3298, JapanSection of Environmental Parasitology, Faculty of Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanWest African Centre for Cell Biology and Infectious Pathogens (WACCBIP), Department of Biochemistry, Cell and Molecular Biology (DBCMB), University of Ghana, Legon, Ghana.Noguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences, University of Ghana, P.O.Box LG 581, Legon, Ghana.