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In vivo and in vitro study of activity of isolated compounds from Brazilian propolis on modulation of neutrophils migration in the inflammatory process

Abstract

The extensive neutrophil migration and the consequent tissue damage are present in several inflammatory diseases. Therefore, the investigation for drugs that interfere with the traffic of neutrophils in the inflammatory process has been increasing. Initial studies of our research group with neovestitol and vestitol, compounds isolated from propolis type 13 (red), and a bioactive fraction identified of geopropolis of Melipona scutellaris showed promising inhibition of neutrophil migration. However, more studies are necessary for the isolation and identification of the compound responsible by the activity of bioactive fraction, as well as demonstrate the efficiency of these isolated compounds on inhibition of migration neutrophils, followed by elucidation of their mechanisms of action. The objective of this study is to evaluate the activity and possible mechanisms of compounds isolated from Brazilian propolis on modulation of neutrophil migration in inflammatory process. The study will be conducted in two phases: the first will be made by the isolation of bioactive compounds (neovestitol and vestitol) of propolis, as well as the isolation and identification of bioactive compound of geopropolis fraction; in the second phase, will be applied biological assays with the isolated compounds of both propolis, where the activity will be evaluated on: migration of neutrophils induced by LPS or mBSA; leukocyte rolling and adhesion in venular endothelium; synthesis and/or liberation of inflammatory mediators (TNF-±, CXCL1/KC, CXCL2 e LTB4) in vivo; expression of molecules like P-selectin, ICAM-1 (present at endothelial cells); synthesis and/or liberation of inflammatory mediators by neutrophils production; the intracellular signaling pathways and the expression of Toll-like receptor (TLR) 2 and TLR4 and B2-integrin in neutrophils in vitro; neutrophils chemotaxis in vitro and the intracellular levels of calcium; the molecular mechanisms of the action and toxicological effects in genomic expression of human cells. It is expected that this study obtains bioactive compounds isolated from two varieties of Brazilian propolis capable of inhibiting neutrophil migration induced by different inflammatory stimuli and has its mechanism of action elucidated. Furthermore, these new information will contribute for the use of these compounds as a novel anti-inflammatory agents or prototypes of new drugs. (AU)

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Scientific publications (6)
(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)
NANI, BRUNO DIAS; FRANCHIN, MARCELO; LAZARINI, JOSY GOLDONI; FREIRES, IRLAN ALMEIDA; DA CUNHA, MARCOS GUILHERME; BUENO-SILVA, BRUNO; DE ALENCAR, SEVERINO MATIAS; MURATA, RAMIRO MENDONCA; ROSALEN, PEDRO LUIZ. Isoflavonoids from Brazilian red propolis down-regulate the expression of cancer-related target proteins: A pharmacogenomic analysis. Phytotherapy Research, v. 32, n. 4, p. 750-754, APR 2018. Web of Science Citations: 2.
BUENO-SILVA, BRUNO; FRANCHIN, MARCELO; ALVES, CLAUDINEY DE FREITAS; DENNY, CARINA; COLON, DAVID FERNANDO; CUNHA, THIAGO MATTAR; ALENCAR, SEVERINO MATIAS; NAPIMOGA, MARCELO HENRIQUE; ROSALEN, PEDRO LUIZ. Main pathways of action of Brazilian red propolis on the modulation of neutrophils migration in the inflammatory process. Phytomedicine, v. 23, n. 13, p. 1583-1590, DEC 1 2016. Web of Science Citations: 15.
FRANCHIN, MARCELO; COLON, DAVID F.; DA CUNHA, MARCOS G.; CASTANHEIRA, FERNANDA V. S.; SARAIVA, ANDRE L. L.; BUENO-SILVA, BRUNO; ALENCAR, SEVERINO M.; CUNHA, THIAGO M.; ROSALEN, PEDRO L. Neovestitol, an isoflavonoid isolated from Brazilian red propolis, reduces acute and chronic inflammation: involvement of nitric oxide and IL-6. SCIENTIFIC REPORTS, v. 6, NOV 7 2016. Web of Science Citations: 9.
DA CUNHA, MARCOS GUILHERME; FRANCHIN, MARCELO; DE PAULA-EDUARDO, LAILA FACIN; FREIRES, IRLAN ALMEIDA; BEUTLER, JOHN A.; DE ALENCAR, SEVERINO MATIAS; IKEGAKI, MASAHARU; MACHADO TABCHOURY, CINTHIA PEREIRA; CUNHA, THIAGO MATTAR; ROSALEN, PEDRO LUIZ. Anti-inflammatory and anti-biofilm properties of ent-nemorosone from Brazilian geopropolis. Journal of Functional Foods, v. 26, p. 27-35, OCT 2016. Web of Science Citations: 2.
FRANCHIN, MARCELO; ROSALEN, PEDRO LUIZ; DA CUNHA, MARCOS GUILHERME; SILVA, RANGEL LEAL; COLON, DAVID F.; BASSI, GABRIEL SHIMIZU; DE ALENCAR, SEVERINO MATIAS; IKEGAKI, MASAHARU; ALVES-FILHO, JOSE C.; CUNHA, FERNANDO Q.; BEUTLER, JOHN A.; CUNHA, THIAGO MATTAR. Cinnamoyloxy-mammeisin Isolated from Geopropolis Attenuates Inflammatory Process by Inhibiting Cytokine Production: Involvement of MAPK, AP-1, and NF-kappa B. Journal of Natural Products, v. 79, n. 7, p. 1828-1833, JUL 2016. Web of Science Citations: 9.
FRANCHIN, MARCELO; COLON, DAVID F.; CASTANHEIRA, FERNANDA V. S.; DA CUNHA, MARCOS G.; BUENO-SILVA, BRUNO; ALENCAR, SEVERINO M.; CUNHA, THIAGO M.; ROSALEN, PEDRO L. Vestitol Isolated from Brazilian Red Propolis Inhibits Neutrophils Migration in the Inflammatory Process: Elucidation of the Mechanism of Action. Journal of Natural Products, v. 79, n. 4, p. 954-960, APR 2016. Web of Science Citations: 13.

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Filed patent(s) as a result of this research project

COMPOSIÇÃO FARMACÊUTICA E SEUS USOS BR1020160262569 - Universidade Estadual de Campinas (UNICAMP) ; Universidade Federal de Alfenas (UNIFAL) . Pedro Luiz Rosalen; Marcos Guilherme Da Cunha; Marcelo Franchin; Masaharu Ikegaki - November 2016, 07

Composição farmacêutica e seus usos PCT/BR2017/000051 - National Institutes of Health ; Universidade de São Paulo (USP) ; Universidade Estadual de Campinas (UNICAMP) ; Universidade Federal de Alfenas (UNIFAL) . Pedro Luiz Rosalen; Masaharu Ikegaki; John Beutler; Thiago Mattar Cunha; Severino Matias de Alencar; Marcos Guilherme da Cunha; Marcelo Franchin - May 2017, 16