Carajurin Induces Apoptosis in Leishmania amazonensis Promastigotes through Reactive Oxygen Species Production and Mitochondrial Dysfunction

Abstract

Carajurin is the main constituent of Arrabidaea chica species with reported anti-Leishmania activity. However, its mechanism of action has not been described. This study investigated the mechanisms of action of carajurin against promastigote forms of Leishmania amazonensis. Carajurin was effective against promastigotes with IC50 of 7.96 ± 1.23 μg.mL−1 (26.4 µM), and the cytotoxic concentration for peritoneal macrophages was 258.2 ± 1.20 μg.mL−1 (856.9 µM) after 24 h of treatment. Ultrastructural evaluation highlighted pronounced swelling of the kinetoplast with loss of electron-density in L. amazonensis promastigotes induced by carajurin treatment. It was observed that carajurin leads to a decrease in the mitochondrial membrane potential (p = 0.0286), an increase in reactive oxygen species production (p = 0.0286), and cell death by late apoptosis (p = 0.0095) in parasites. Pretreatment with the antioxidant NAC prevented ROS production and significantly reduced carajurin-induced cell death. The electrochemical and density functional theory (DFT) data contributed to support the molecular mechanism of action of carajurin associated with the ROS generation, for which it is possible to observe a correlation between the LUMO energy and the electroactivity of carajurin in the presence of molecular oxygen. All these results suggest that carajurin targets the mitochondria in L. amazonensis. In addition, when assessed for its drug-likeness, carajurin follows Lipinski’’s rule of five, and the Ghose, Veber, Egan, and Muegge criteria.

Direct In Vitro Comparison of the Anti-Leishmanial Activity of Different Olive Oil Total Polyphenolic Fractions and Assessment of Their Combined Effects with Miltefosine

“…promastigotes and provoked the generation of intracellular ROS [ 28 ]. These findings are consistent with other studies which also demonstrated the contribution of ROS production and mitochondrial dysfunction to Leishmania death induced by natural products, such as sesamol, carajurin, apigenin, and quercetin [ 37 , 38 , 39 , 40 ].…”

Holanamine, a Steroidal Alkaloid from the Bark of Holarrhena pubescens Wall. ex G. Don Inhibits the Growth of Leishmania donovani by Targeting DNA Topoisomerase 1B

“…ROS are highly reactive and modify the proteins, lipids, and nucleic acids. Therefore, ROS-mediated cell damage is a frequent event in apoptosis. ,, Our results, indicating that holanamine causes oxidative stress by inducing ROS generation, are supported by prior research showing that a number of natural products trigger ROS formation inside parasites, which has detrimental effects on them …”

“…A cascade of biochemical and physical changes occurs in the cell during apoptosis. Plant secondary metabolites such as gallic acid, apigenin, ferulic acid, flavonoids, and alkaloids have been reported to have antileishmanial activity by triggering apoptosis in Leishmania parasites . We have also noticed a similar apoptotic process in holanamine-treated L.…”

In silico-based screening for natural product’s structural analogs as new drugs candidate against leishmaniasis

“…Recently, the anthocyanidin profile of Arrabidaea chica has been examined and their antileishmanial activity was analyzed [73]; whereas Carajurin (PubChem CID: 44257040) showed the highest activity against the intracellular parasites, altering all parameters of in vitro infection [74]. Additionally, it has been shown that Carajurin leads to a decrease in the mitochondrial membrane potential, an increase in reactive oxygen species production, and cell death by late apoptosis in L. amazonensis [75].…”