Evaluation of Anti- Activity of Chemical Constituents from Isolated Using High-Performance Countercurrent Chromatography.

Endemic in 21 countries, Chagas disease, also known as American Trypanosomiasis, is a neglected tropical disease (NTD) caused by the protozoan parasite . The available drugs for the treatment of this disease, benznidazole and nifurtimox, are outdated and display severe side effects. Thus, the discovery of new drugs is crucial. Based on our continuous studies aiming towards the discovery of natural products with anti- potential, the MeOH extract from aerial parts of Dusén ex. Malme (Asteraceae) displayed activity against this parasite and was subjected to high-performance countercurrent chromatography (HPCCC), to obtain one unreported -labdane diterpene - sphenophyllol () - as well as the known compounds gaudichaudol C (), -kaurenoic acid (), hispidulin (), eupafolin (), and one mixture of di--caffeoylquinic acids (-). Compounds - were characterized by analysis of nuclear magnetic resonance (NMR) and mass spectrometry (MS) data. When tested against trypomastigote forms, isolated labdane diterpenes and displayed potent activity, with EC values of 20.1 μM and 2.9 μM, respectively. The mixture of chlorogenic acids -, as well as the isolated flavones and , showed significant activity against the clinically relevant amastigotes, with EC values of 24.9, 12.8, and 2.7 μM, respectively. Nonetheless, tested compounds - displayed no cytotoxicity against mammalian cells (CC > 200 μM). These results demonstrate the application of HPCCC as an important tool to isolate bioactive compounds from natural sources, including the antitrypanosomal extract from , allowing for the development of novel strategic molecular prototypes against tropical neglected diseases.