Aspidosperma species as sources of anti-malarials: uleine is the major anti-malarial indole alkaloid from Aspidosperma parvifolium (Apocynaceae).

Maria Fâni Dolabela,Fabíola Dutra Rocha,Luciana Ferreira Soares,Alaíde Braga De Oliveira,Marinete Marins Póvoa,Renata Cristina De Paula,Geraldo Célio Brandão

doi:10.1186/s12936-015-0997-4

Abstract

BackgroundSeveral species of the genus Aspidosperma (Apocynaceae) are used for the treatment of human malaria in Brazil and other meso- and South American countries.MethodsEthanol extract from Aspidosperma parvifolium trunk bark was submitted to acid–base extractions leading to alkaloid and neutral fractions. The alkaloid fraction was chromatographed over a silica gel column. Ethanol extract, fractions and uleine were analysed by HPLC–DAD, UPLC-ESI–MS/MS and HPLC-ESI-MicroTOF-MS. The anti-malarial activity was assayed against resistant and sensitive chloroquine Plasmodium falciparum strains by microscopic, [3H]-hypoxanthine incorporation and HRPII techniques. Cytotoxicity (CC50) was evaluated against Vero and HepG2 cell lines by the MTT technique; selectivity indexes (SI = CC50/IC50) were calculated.ResultsThe major peak in the HPLC–DAD chromatograms of the ethanol extract, alkaloid and neutral fractions suggested the presence of uleine that was isolated from the alkaloid fraction by column chromatography and was characterized by spectroscopic methods. A total of 15 alkaloids, besides uleine, were identified in the alkaloid fraction by UPLC-DAD-ESI–MS/MS and HPLC-ESI-MicroTOF-MS. The ethanol extract from Aspidosperma parvifolium and the neutral fraction were moderately active against P. falciparum strains. The alkaloid fraction and uleine disclosed high anti-malarial activity against chloroquine-resistant P. falciparum strain (IC50 < 1 µg/mL). The ethanol extract, neutral fraction and uleine showed low cytotoxicity against Vero and HepG2 cell lines (CC50 > 300 µg/mL). The alkaloid fraction showed moderate cytotoxicity to HepG2 cell line (CC50 = 74.4 µg/mL). High SI values (>10) were determined for all samples.ConclusionEthanol extract from Aspidosperma parvifolium trunk bark afforded uleine that is the major constituent of the alkaloid fraction and disclosed a good in vitro anti-malarial activity. Moreover, 15 other indole alkaloids have been identified along with uleine.

Highlights

Several species of the genus Aspidosperma (Apocynaceae) are used for the treatment of human malaria in Brazil and other meso- and South American countries
The present paper reports on the anti-malarial activity of Aspidosperma parvifolium against chloroquine-resistant (W2) and sensitive (3D7) P. falciparum strains
Phytochemical studies HPLC–DAD chromatograms showed a peak for the same major component in the ethanol extract (RT = 30.1 min; Fig. 1a), alkaloid fraction (RT = 29.6 min, Fig. 1b) and neutral fraction (RT = 29.8 min, Fig. 1c) that might correspond to uleine (Fig. 1d)

Summary

Introduction

Several species of the genus Aspidosperma (Apocynaceae) are used for the treatment of human malaria in Brazil and other meso- and South American countries. The extensive use of antimalarial drugs has imposed a high selective pressure on parasites, leading to the emergence of drug resistance, in Plasmodium falciparum [2]. Plasmodium falciparum resistance to artemisinins has been detected in four countries of the Southeast Asia region. Artemisinin derivatives comprise the therapeutic scheme recommended for malaria treatment [3]. This points to the urgency for the development of new anti-malarial drugs. Plants continue to be a valuable source of bioactive compounds and investigation of traditional medicines used to treat malaria in endemic countries has afforded useful anti-malarial drugs, such as quinine, artemisinins and atovaquone [4,5,6]

Methods

Results

Conclusion