Organic Propolis is a singular type of propolis, since it is produced under organic and certified conditions in Atlantic Brazilian forest in the states of Paraná and Santa Catarina. In a previous study, we chemically evaluated (TLC and HPLC) the OP hydroethanolic extracts (obtained from 14 organic beekeepers) and we have demonstrated that: OP can be grouped in 7 chemical profiles; presents unique compounds; and can be considered as a new typo of Brazilian propolis, because many of these compounds cannot be found in other Brazilian propolis types. We have also showed that OP has great antioxidant activity and that the profile 6 (OP) has anti-inflammatory activity because it can inhibit the NF:B transcription factor and the TNF-± release. These findings are a strong evidence that shows that OP6 can inhibit the neutrophil migration, phenomenon that is considered the ideal target for treatment of inflammatory diseases without effective treatment, such as the periodontal disease. Our results were even more evidenced by a pharmacogenomics study, which showed that OP6 is able to reduce the expression of genes enrolled with the leukocyte migration. Therefore, this study aim to isolate and to identify the OP6 compounds with inhibitory activity on neutrophil migration in vitro, in vivo and in silico, as well to elucidate their mechanism of action. The OP samples will be collected from the same beekeepers which provided the samples used in the previous work and these samples will be extract with hydroethanolic solution. The extracts will be evaluated by TLC and only those similar to the OP6 profile will be selected and submitted to a prospection bioguided by its effects in inhibition of neutrophil migration in vitro. The bioprospection will be realized by three chemical fractionation processes: diol silica, reverse phase silica and LH20 sephadex gel. In each fractionation step, we will obtain fractions or subfractions of OP6, which will be submitted to a chemical (TLC) and biological monitoring by the in vitro experiments for evaluation of cell viability, inflammatory citokines release quantification, neutrophil chemotaxis and adhesion proteins expression. The fractions or subfractions with greater yield and biological activity will be selected and, in the end of this processes, the molecules responsible for the effects will be identified thought semi-preparative HPLC, nuclear magnetic resonance and high resolution mass spectrometry. Posteriorly, these molecules will be submitted to an in silico experiment in which we will obtain its pharmacokinetic and toxicological properties; and in vivo (C57BL/6 mice) to confirm its inhibitory properties in neutrophil migration processes with the experiments of peritonitis, intravital microscopy, cytokine and chemokine release quantification and adhesion proteins expression. The study will be finished with the confirmation of the molecular mechanisms of action of OP6 compounds by a pharmacogenomics study. Hence, we expect to isolate and to identify the OP6 compounds with inhibitory effects in the neutrophil migration, as well to elucidate its molecular mechanisms and that these molecules can be effective in the treatment of inflammatory diseases.
News published in Agência FAPESP Newsletter about the scholarship: