Síntese e perfil anti-inflamatório in vitro e in vivo de derivados fenilacetamida e fenilacrilamida

Abstract

Therapeutic approaches to the treatment of inflammatory conditions are focused on suppressing, blocking or inhibiting inflammatory mediators. The drugs available on the market for these disorders, although effective, have as limitations gastrointestinal, renal, cardiovascular and metabolic disorders, making the development of new drugs necessary. The objective of this study was to develop, characterize and evaluate the in vitro and in vivo anti-inflammatory profile of the derivatives: 2-cyan N-phenylacetamide - JM-01; (E) -2-cyano-3- (1H-indol-3-yl) - N-phenylacrylamide - ICMD-01 and (E) -2-cyano-N,3-diphenylacrylamide -JMPR-01 and later determine the toxicological potential of the most promising compound. The compounds were obtained through an amidation reaction between substituted anilines and ethyl 2-cyanoacetate, followed by a Knoevenagel-type condensation reaction with substituted aldehydes, the structural elucidation being performed by spectroscopic and spectrometric techniques. To evaluate the anti-inflammatory potential in vitro, cell viability tests, cytokine and nitrite dosages in J774 macrophage cultures and in vivo: zymosan-induced peritonitis, edema, inflammatory hyperalgia, cytokine and prostaglandin E2 measurements CFA-induced. The antinociceptive effect and motor adaptive changes of animals treated with the compounds were evaluated by tail-flick and rota rod tests. The determination of safety of use was performed using single dose acute toxicity models, in vitro hemolytic activity and ulcerogenic susceptibility. Pharmacokinetic predictions were performed using the SwissADME and Xenosite platforms. The synthesis of the compounds JM-01, JMPR-01 and ICMD-01 proved feasible, with satisfactory yields easy to purify and with spectrometric and spectroscopic results consistent with the structure. In vitro results for the compounds showed significant inhibitions (p<0.05) of nitrite and cytokine production in concentrations up to 50 μM for JM-01 and JMPR-01 and 100 μM for ICMD-01. In the zymosan-induced peritonitis test all the derivatives showed inhibition of leukocyte migration at doses of (5, 10 and 50 mg kg^-1) significantly different from the control group (p<0.05) with activity higher than the group treated with indomethacin at doses of 10 mg kg^-1.In CFA-induced paw edema the compound with the best profile of antiedematogenic activity in times from 2 to 6 hours was the ICMD-01 (50 mg kg^-1), presenting respectively inhibition of 44.45, 57.14 and 67.34%. The ICMD-01 also showed a significant reduction (p<0.05) of local IL-1β and TNF-α and PGE2 levels when compared to the control group. For IL-1β and PGE2 inhibition promoted by the ICMD-01 is comparable to that of dexamethasone (2mg kg^-1), while for TNF-α the compound proved superior. Regarding the sedative, antinociceptive and central activity none of the compounds induced considerable effects. In the toxicological profile of ICMD-01, no relevant clinical signs or death were evidenced during the acute toxicity experiment, besides a good gastric tolerance. Also no excess degree of hemolytic activity was evidenced. In the pharmacokinetic predictions it was observed that the compounds JM-01, JMPR-01 and ICMD-01, are potentially well absorbed by the TGI and permeable to Hb in addition, have points susceptible to metabolization that can influence the pharmacological effect. These initial studies contribute to validate the biological potential of these derivatives, making them strong candidates for antiinflammatory drugs with low ulcerogenic potential.