Antiproliferative activity and new argininyl bufadienolide esters from the "cururu" toad Rhinella (Bufo) schneideri

Abstract

Ethnopharmacological relevance: Toads known as "cururu" (Rhinella schneideri) have been used in the Brazilian Pantanal and Paraguayan Chaco wetlands to treat erysipelas and cancer. The aim of the study was to assess the antiproliferative effect of the venom obtained from Rhinella schneideri and to identify its constituents by spectroscopic and spectrometric methods.

Materials and methods: The venom was obtained by gentle pressing the parotid glands of the toads. The dry crude drug was analyzed by HPLC-MS-MS and chromatographed on Sephadex LH-20 to obtain purified compounds and fractions for spectroscopic analysis. The venom and fractions were evaluated for antiproliferative activity towards normal human lung fibroblasts (MRC-5) and four human cancer cell lines: gastric epithelial adenocarcinoma (AGS), lung cancer (SK-MES-1), bladder carcinoma (J82) and promyelocytic leukemia (HL-60).

Results: From the Rhinella schneideri venom, 29 compounds were isolated and/or identified by spectroscopic and spectrometric means. Three known alkaloids and five argininyl diacids were identified in the complex mixture by HPLC-MS-MS. Nine out of fifteen argininyl diacid derivatives of the bufadienolides bufalin, marinobufagin and telocinobufagin are reported for the first time and four argininyl diacids are described for the first time as natural products. The venom and the fractions 9-13 showed a remarkable antiproliferative effect, with IC50 values in the range 0.019-0.022, 0.035-0.040, 0.028-0.064, 0.042-0.056 and 0.044-0.052 mu g/mL for MRC-5, AGS, SK-MES-1, J82 and HL-60 cell lines, respectively. Under the same experimental conditions, IC50 values of the reference compound etoposide were 2296, 0.277, 1.295, 1.884 and 1.059 mu g/mL towards MRC-5, AGS, SK-MES-1, J82 and HL-60 cells, respectively.

Conclusions: The venom showed a strong antiproliferative effect towards human cancer cells and presented a high chemical diversity in its constituents, supporting its use as anticancer agent. These findings encourage further work on the chemistry and bioactivity of South American toad venoms. (C) 2014 Elsevier Ireland Ltd. All rights reserved.