Leishmaniasis: from screening to the study of mechanisms of action, a contribution to the discovery of new bioactive molecules

• Grant number: 17/03552-5
• Support Opportunities: Regular Research Grants
• Start date: May 01, 2017
• End date: October 31, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Marcia Aparecida Silva Graminha
• Grantee: Marcia Aparecida Silva Graminha
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Leishmaniasis is a pathogenic disease found throughout the world caused by protozoan parasites of the Leishmania genus. Despite the numerous problems associated with existing treatments, pharmaceutical companies continue to neglect the development of better ones. The high toxicity of current drugs combined with emerging resistance to them makes providing new therapeutic alternatives urgent. We have been studying two new antileishmanial molecules: a binuclear cyclopalladated complex containing Pd(II) and N,N'-dimethylbenzylamine [Pd(dmba)(µ-N3)]2,, CP2, against Leishmania amazonensis and a furoxan derivative (E)-4-(4-((2-(3-hydroxybenzoyl)hydrazone)methyl)phenoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide, 14e.. Preliminary results showed that CP2 inhibits both promastigote (IC50 = 13.15 ± 0.67) and intracellular amastigote proliferation (IC50 = 10.14 ± 2.20), with high selectivity to the parasite. In vivo assays using L. amazonensis-infected BALB/c showed an 80% reduction in the parasite load when compared to infected and un-treated animals. Also, compared to amphotericin B treatment, CP2 did not show any side effects, which was corroborated by the analysis of plasma levels of different hepatic and renal biomarkers. Furthermore, CP2 was able to inhibit the Leishmania donovani Topoisomerase 1B (Ldtopo1B), one of the most important targets in Kinetoplastida. The furoxan 14e has shown good antileishmanial activity (EC50 = 3.14 µM) against intracellular amastigote forms of L. (L.) infantum and high selectivity to the parasite (SI = 66.35) was 2.8-times superior to that presented by amphotericin B (AmpB). Treatment of L. (L.) infantum-infected hamsters with compound 14e at 3.00 mg/Kg/day led to a meaningful reduction of parasite load in spleen (49.87%) and liver (54.24%), respectively; these data were corroborated by histopathological analysis, which also revealed reduction in the number of inflammatory cells in the liver and the spleen of the treated animals. The parasitemia reduction might be related to NO, since this molecule is a NO-donor. We did not observe neither side effects nor elevation of hepatic/ renal biomarkers in the plasma levels. Based on these data, we decided to further investigate the potential mechanism of action of these two molecules. For CP2, we are going to validate the proteomic data obtained during the development of the research project FAPESP number 2013/08248-1 as well as investigate if CP2 is interfering with calcium homoestasis. For 14e, proteomic analysis and validation will be made. In parallel, our research group will be doing screening for antileishmanial activity of synthetic or natural compounds. (AU)

Scientific publications

(15)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DO ESPIRITO SANTO, RAFAEL DIAS; ARENAS VELASQUEZ, ANGELA MARIA; GUSHIKEN PASSIANOTO, LUANA VITORINO; LOPERA SEPULVEDA, ALEX ARBEY; CLEMENTINO, LEANDRO DA COSTA; ASSIS, RENATA PIRES; BAVIERA, AMANDA MARTINS; KALABA, PREDRAG; DOS SANTOS, FABIO NEVES; EBERLIN, MARCOS NOGUEIRA; et al. N, N `, N `'-trisubstituted guanidines: Synthesis, characterization and evaluation of their leishmanicidal activity. EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, v. 171, p. 116-128, JUN 1 2019. (13/24487-6, 17/03552-5, 16/19289-9, 13/08248-1)

LINARES, IRWIN A. P.; VELASQUEZ, ANGELA M. A.; GRAMINHA, MARCIA A. S.; DE OLIVEIRA, KLEBER T.; PERUSSI, JANICE RODRIGUES. Antileishmanial activity of amphiphilic chlorin derivatives mediated by photodynamic therapy. Photodiagnosis and Photodynamic Therapy, v. 31, p. 9-pg., 2020-09-01. (18/00106-7, 17/03552-5, 13/07276-1, 19/21661-1)

CLEMENTINO, LEANDRO DA COSTA; ODA, FERNANDO BOMBARDA; TEIXEIRA, THAIZ RODRIGUES; TAVARES, RENATA SPAGOLLA NAPOLEAO; COLEPICOLO, PIO; DOS SANTOS, ANDRE GONZAGA; DEBONSI, HOSANA MARIA; GRAMINHA, MARCIA A. S.. The antileishmanial activity of the antarctic brown algaAscoseira mirabilisSkottsberg. NATURAL PRODUCT RESEARCH, v. 35, n. 23, p. 5-pg., 2020-06-19. (17/03552-5)

MOREIRA, VITOR PARTITE; DA SILVA MELA, MICHELE FERREIRA; DOS ANJOS, LUANA RIBEIRO; SARAIVA, LEONARDO FIGUEIREDO; VELASQUEZ, ANGELA M. ARENAS; KALABA, PREDRAG; FABISIKOVA, ANNA; CLEMENTINO, LEANDRO DA COSTA; AUFY, MOHAMMED; STUDENIK, CHRISTIAN; et al. Novel Selective and Low-Toxic Inhibitor of LmCPB2.8 Delta CTE (CPB) One Important Cysteine Protease for Leishmania Virulence. BIOMOLECULES, v. 12, n. 12, p. 21-pg., 2022-12-01. (18/00581-7, 20/04415-4, 16/19289-9, 17/03552-5, 18/23015-7, 21/02595-8)

CLEMENTINO, LEANDRO DA COSTA; ARENAS VELASQUEZ, ANGELA MARIA; PASSALACQUA, THAIS GABAN; DE ALMEIDA, LETICIA; GRAMINHA, MARCIA A. S.; MARTINS, GILMARCIO Z.; SALGUEIRO, LIGIA; CAVALEIRO, CARLOS; SOUSA, MARIA DO CEU; MOREIRA, RAQUEL R. D.. In vitro activities of glycoalkaloids from the Solanum lycocarpum against Leishmania infantum. REVISTA BRASILEIRA DE FARMACOGNOSIA-BRAZILIAN JOURNAL OF PHARMACOGNOSY, v. 28, n. 6, p. 673-677, NOV-DEC 2018. (17/03552-5)

DE ANNUNZIO, SARAH RAQUEL; SILVA COSTA, NATALIA CAROLINE; MEZZINA, RAFAELA DALBELLO; GRAMINHA, MARCIA A. S.; FONTANA, CARLA RAQUEL. Chlorin, Phthalocyanine, and Porphyrin Types Derivatives in Phototreatment of Cutaneous Manifestations: A Review. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 20, n. 16, AUG 2 2019. (18/23015-7, 17/03552-5, 18/09088-1)

TEIXEIRA, THAIZ RODRIGUES; RANGEL, KAREN CRISTINA; NAPOLEAO TAVARES, RENATA SPAGOLLA; KAWAKAMI, CAMILA MARTINS; DOS SANTOS, GUSTAVO SOUZA; MARIA-ENGLER, SILVYA STUCHI; COLEPICOLO, PIO; GASPAR, LORENA RIGO; DEBONSI, HOSANA MARIA. In Vitro Evaluation of the Photoprotective Potential of Quinolinic Alkaloids Isolated from the Antarctic Marine Fungus Penicillium echinulatum for Topical Use. MARINE BIOTECHNOLOGY, v. 23, n. 3, p. 357-372, JUN 2021. (14/50926-0, 17/03552-5)

DE ALMEIDA, LETICIA; PASSALACQUA, THAIS GABAN; DUTRA, LUIZ ANTONIO; VARONEZ DA FONSECA, JESSICA N.; QUEIROZ NASCIMENTO, RHAYANNE F.; IMAMURA, KELY BRAGA; DE ANDRADE, CLEVERTON ROBERTO; DOS SANTOS, JEAN LEANDRO; GRAMINHA, MARCIA A. S.. In vivo antileishmanial activity and histopathological evaluation in Leishmania infantum infected hamsters after treatment with a furoxan derivative. BIOMEDICINE & PHARMACOTHERAPY, v. 95, p. 536-547, NOV 2017. (17/03552-5, 13/08248-1)

RANGEL, KAREN C.; DEBONSI, HOSANA M.; CLEMENTINO, LEANDRO C.; GRAMINHA, MARCIA A. S.; VILELA, LEONARDO Z.; COLEPICOLO, PIO; GASPAR, LORENA R.. Antileishmanial activity of the Antarctic red algae Iridaea cordata (Gigartinaceae; Rhodophyta). JOURNAL OF APPLIED PHYCOLOGY, v. 31, n. 2, p. 825-834, APR 2019. (17/03552-5, 16/06931-4)

CLEMENTINO, LEANDRO DA COSTA; ODA, FERNANDO BOMBARDA; TEIXEIRA, THAIZ RODRIGUES; TAVARES, RENATA SPAGOLLA NAPOLEAO; COLEPICOLO, PIO; DOS SANTOS, ANDRE GONZAGA; DEBONSI, HOSANA MARIA; GRAMINHA, MARCIA A. S.. The antileishmanial activity of the antarctic brown algaAscoseira mirabilisSkottsberg. NATURAL PRODUCT RESEARCH, JUN 2020. (17/03552-5)

LOPERA SEPULVEDA, ALEX ARBEY; ARENAS VELASQUEZ, ANGELA MARIA; PATINO LINARES, IRWIN ALEXANDER; DE ALMEIDA, LETICIA; FONTANA, CARLA RAQUEL; GARCIA, CLAUDIA; SILVA GRAMINHA, MARCIA APARECIDA. Efficacy of photodynamic therapy using TiO2 nanoparticles doped with Zn and hypericin in the treatment of cutaneous Leishmaniasis caused by Leishmania amazonensis. Photodiagnosis and Photodynamic Therapy, v. 30, JUN 2020. (16/05345-4, 17/03552-5, 16/19289-9, 18/23015-7)

LOPERA, A. A.; VELASQUEZ, A. M. A.; CLEMENTINO, L. C.; ROBLEDO, S.; MONTOYA, A.; DE FREITAS, L. M.; BEZZON, V. D. N.; FONTANA, C. R.; GARCIA, C.; GRAMINHA, M. A. S.. Solution-combustion synthesis of doped TiO2 compounds and its potential antileishmanial activity mediated by photodynamic therapy. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY, v. 183, p. 64-74, JUN 2018. (17/03552-5, 16/05345-4, 16/19289-9, 14/24581-5, 13/08248-1)

VELASQUEZ, ANGELA M. A.; BARTLETT, PAULA J.; LINARES, IRWIN A. P.; PASSALACQUA, THAIS G.; TEODORO, DAPHNE D. L.; IMAMURA, KELY B.; VIRGILIO, STELA; TOSI, LUIZ R. O.; LEITE, ALINE DE LIMA; BUZALAF, MARILIA A. R.; et al. ew Insights into the Mechanism of Action of the Cyclopalladated Complex (CP2) in Leishmania: Calcium Dysregulation, Mitochondrial Dysfunction, and Cell Deat. Antimicrobial Agents and Chemotherapy, v. 66, n. 1, JAN 2022. (16/18191-5, 16/05345-4, 18/23015-7, 20/04415-4, 17/03552-5, 16/19289-9, 19/21661-1)

CLEMENTINO, LEANDRO DA COSTA; SANTOS FERNANDES, GUILHERME FELIPE; PROKOPCZYK, IGOR MUCCILO; LAURINDO, WILQUER CASTRO; TOYAMA, DANYELLE; MOTTA, BRUNO PEREIRA; BAVIERA, AMANDA MARTINS; HENRIQUE-SILVA, FLAVIO; DOS SANTOS, JEAN LEANDRO; GRAMINHA, MARCIA A. S.. esign, synthesis and biological evaluation of N-oxide derivatives with potent in vivo antileishmanial activit. PLoS One, v. 16, n. 11, NOV 1 2021. (16/06931-4, 17/03552-5)

DO ESPIRITO SANTO, RAFAEL DIAS; ARENAS VELASQUEZ, ANGELA MARIA; GUSHIKEN PASSIANOTO, LUANA VITORINO; LOPERA SEPULVEDA, ALEX ARBEY; CLEMENTINO, LEANDRO DA COSTA; ASSIS, RENATA PIRES; BAVIERA, AMANDA MARTINS; KALABA, PREDRAG; DOS SANTOS, FABIO NEVES; EBERLIN, MARCOS NOGUEIRA; et al. N, N ', N ''-trisubstituted guanidines: Synthesis, characterization and evaluation of their leishmanicidal activity. EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, v. 171, p. 13-pg., 2019-06-01. (13/24487-6, 17/03552-5, 13/08248-1, 16/19289-9)