Certain species of plants are popularly used to treat accidents with venomous animals and their studies demonstrate interest from the phytochemical point of view, thus contributing to the isolation and identification of biologically active compounds. The genus Bredemeyera, belonging to the Polygalaceae family, comprises species popularly known as "snake root" due to its use as an antivenom. In ad dition to their antivenom properties, they are also known for their expectorant, diuretic and hypotensive properties. The chromatographic study of Bredemeyra floribunda resulted in the isolation of two secondary metabolites, the flavonoid rutin and xanthone, called 1,7-dihydroxy-3,4,8- trimethoxyxanthone and a fraction containing a mixture of two saponins. In view of all this, the objective of this study was to investigate the phytochemical profile of the hydroethanolic extract of the roots of B. floribunda using dereplication through ultra-performance liquid chromatography coupled with mass spectrometry (CLUE-MS) and to evaluate its effects of extract and isolated substances against the main enzymatic  activities  of  the  venom  of  the  snake  Bhotrops  leucurus  in  vitro.  The  CLUE-MS  analysis  of  the  hydroethanolic  extract  of Bredemeyera floribunda roots revealed that flavonoids, xanthones and saponins constitute the most relevant class of secondary metabolites in the extracts. For the enzymatic inhibition studies, extracts and previously isolated substances were tested, for the proteolytic assay it was observed that the proportions 1:25, 1:50, 1:75 and 1:100 (poison:extract, w/w) the extract significantly reduced enzymatic activity. The same way was evidenced in the phospholipase activity, all proportions of the tested extract proved to be effective in inhibiting the enzymatic effect. In the hyaluronidase method, it was demonstrated that the extract was effective in ratios 1:10 and 1:25. These data revealed the antiophidic potential of the extracts by inhibiting the main enzymatic activities of the B. leucurus snake venom, demonstrating that both extracts can be relevant and promising for the treatment of snake bites.