Avaliação da atividade anticoagulante e bloqueadora de elastase de um inibidor de tripsina isolado de Caesalpinea pyramidalis Tul.

Abstract

Plants are subject to a diversity of unfavorable biotic and abiotic conditions, undergoing constantly some evolutionary adaptation, that can contribute to its reproduction and survival. The development of sophisticated defense strategies against pests and pathogens such as the synthesis of several natural bioactive compounds stands out as part of this adaptive biological arsenal. A key group of bioactive molecules in this context are the protease inhibitors, which compromise the activity of digestive enzymes of herbivores, as well as an action of pathogens, contributing to the balance of biological interactions. Protease inhibitors, are important molecules because of their heterologous potential in other biological systems, and may have in vitro and in vivo effects on several biological models, as well as therapeutic properties in a range of disorders that compromise human health. In this work an inhibitor of serinoprotease (CpTI), was isolated from the Caesalpinia pyramidalis Tul. seeds by fractionation of the crude extract with 30- 60% ammonium sulfate, followed by separation in affinity chromatography on TrypsinSepharose 4B-CNBr-actived column, the retained peak was enriched in inhibitory activity for trypsin, chymotrypsin and elastase serine protease type. The thermostability test showed that CpTI was able to maintain its functionality up to 80 °C and the concentrations required to inhibit 50% of the activity of the trypsin, human neutrophil elastase and chymotrypsin enzymes were 17, 21 and 156 μg/mL, respectively. CpTI also prolonged the coagulation time by the intrinsic pathway (TPPa) in more than 80 seconds at a concentration of 36.2 μg / mL. It was also observed that CpTI has no cytotoxic activity to red blood cells at any of the concentrations tested (0,03 , 0,125 e 0,5 mg/mL). These results demonstrate the potential therapeutic use of CpTI in processes involving inflammation and coagulation.