Cysteine Protease Inhibitors from the Methanol Extract of the Root Bark of Securidaca longepedunculata with Antimalarial Potentials in Chloroquine-Resistant P. berghei Parasite

Malaria parasite resistance against Artemisinin-based Combination Therapy (ACT) in some parts of the world necessitates the search for antimalarial compounds or plant extracts with novel mode of action and active against the chloroquine-resistant strain of the parasite to serve as alternative to ACT. Cysteine protease inhibitors' fraction of the methanol extract of the root bark of Securidaca longepedunculata (CPI) was investigated for antiplasmodial activity against chloroquine-resistant P. berghei-infected mice and its inhibitory effect on papain, P. berghei cysteine proteases and heme biocystallization were also evaluated. The methanol extract of the root bark was obtained by soxhlet extraction with 1,000 mL of 70% (v/v) methanol for 48 hours and concentrated to dryness at 45∘C. CPI was obtained using PBS (pH 7) extraction followed by cold acetone precipitation. Peter's four-day suppressive and Rane's four-day curative test was employed to assess the antimalarial potentials of CPI. Data was analysed with one way ANOVA followed by Dunnett's post hoc test, differences were considered significant at p≤0.05. The suppressive effect of CPI was significant (p≤0.05) at 34 and 23 mg/kg doses. Doses of 34, 23 and 11 mg/kg produced significant (p≤0.05) dose-dependent curative effect. CPI inhibited the proteolytic activity of papain enzyme and P. berghei cysteine proteases in vitro with IC50 values of 20.1 and 5.6 μg/mL respectively. The present study showed that cysteine protease inhibitors fraction of the methanol extract of the root bark of Securidaca longepedunculata is a potential source of novel antimalarial agents that could target malaria parasite cysteine proteases.