Phytochemistry and in vitro pharmacological activities of South African Vitex (Verbenaceae) species

The in vitro phytochemical and pharmacological investigation of the non-volatile extracts of five South African Vitex species (Verbenaceae); V. obovata ssp. obovata, V. obovata ssp. wilmsii, V. pooara, V. rehmannii and V. zeyheri were investigated in order to validate their traditional use to treat a wide range of ailments such as malaria, wounds, skin diseases and body pains. The antimicrobial activity was assessed using the minimum inhibitory concentration assay. Through bioactivity-guided fractionation, the fraction responsible for the antimicrobial activity was determined. The toxicity profile, anti-oxidant and anti-inflammatory activity was evaluated using the tetrazolium cellular viability, 2,2-diphenyl-1-picrylhydrazyl and 5-lipoxygenase assays respectively. The antimalarial activity of the extracts and isolated compound from V. rehmannii was also investigated on the chloroquine-resistant Gambian FCR-3 strain of Plasmodium falciparum using the tritiated hypoxanthine incorporation assay. Mostly good antimicrobial inhibition was evident against Grampositive bacteria (0.02–8.00 mg/ml) and lower activity against the Gramnegative bacteria and the yeast (0.50–8.00 mg/ml). The fraction responsible for antimicrobial activity of V. rehmannii was purified to give a labdane diterpene as an inseparable epimeric mixture of 12 S,16 S/R-dihydroxy- ent-labda-7,13-dien-15,16-olide. Cirsimaritin was also isolated and identified from V. rehmannii. All the species, apart from V. zeyheri, exhibited scavenging activity (IC50: 22.14 ± 1.74 to 33.06 ± 1.68 μg/ml) in the anti-oxidant assay. None of the species displayed any anti-inflammatory activity at 100 μg/ml. All the extracts and the labdane diterpene exhibited good antimalarial activity, with the labdane diterpene being the most active (IC50: 2.39 ± 0.64 μg/ml). The test extracts were shown to be highly toxic, displaying safety index values ranging from 0.53 to 2.59. Of all the pharmacological investigations, the antimalarial and antimicrobial activity exhibited greatest activity and may provide a scientific basis for the ethnomedical use of Vitex species.