Cytological profiling of trypanocidal principles from and .

The use of herbal medicines to treat ailments is a common practice in several regions in Africa, relying on knowledge systems that have evolved over several generations. These herbal remedies are often based on anecdotal claims, many of which lack scientific validation. This study investigates the mode of action of two bioactive fractions, F1 (IC: 8.5 µg/mL) and F5 (IC: 7.4 µg/mL), derived from a dichloromethane extract of a herbal mixture, consisting of and , that is commonly used in Ghana to treat parasitic fevers. Both fractions exhibited trypanocidal effects with minimal cytotoxicity to mammalian cells. F5 induced necrotic cell death through mitochondrial oxidative stress, evidenced by a 3.5-fold increase in mitochondrial reactive oxygen species at 2 × IC (< 0.0001) and significant mitochondrial membrane depolarization (< 0.01). In contrast, F1 primarily disrupted kinetoplast segregation, increasing 2K1 N cells by 3.2-fold at 1 × IC (< 0.0001) and instigating an accumulation of dyskinetoplastic cells (0KXN). Both fractions induced morphological distortions, nuclear fragmentation, and loss of flagellar integrity. This study provides the first mechanistic insights into the antitrypanosomal activity of bioactive fractions obtained from a mixture of and . The distinct targeting of mitochondrial ROS production (F5) and kinetoplast replication (F1) highlights their potential as leads for the development of new antitrypanosomal drugs with novel mechanisms of action. These findings reinforce the value of ethnomedicinal plants as sources of novel bioactive compounds.