Proteomics-driven screening of artemisinin-based combination ratios and mechanistic insights into Plasmodium berghei infection in mice.

Introduction: Malaria, a life-threatening mosquito-borne disease caused by Plasmodium falciparum, poses a substantial health burden on tropical and subtropical regions. Artemisinin, a sesquiterpene lactone isolated from Artemisia annua L., and its derivatives were initially used as monotherapies for malaria treatment. However, limitations such as short pharmacokinetic half-life and emerging drug resistance have driven the widespread adoption of artemisinin-based combination therapies (ACTs) as first-line interventions. A. annua contains other bioactive compounds such as arteannuin B, artemisinic acid, and scopoletin that exhibit distinct pharmacological properties. In this study, we aimed to devise a new strategy for treatment of malaria to overcome artemisinin resistance in Plasmodium species. Methods: We systematically screened antimalarial compound ratios using murine malaria models and optimized a formula comprising arteannuin B, artemisinic acid, and scopoletin. Through integrated proteomic profiling and western blot validation, we elucidated the immunomodulatory mechanisms underlying the antimalarial efficacy of this combination. Results: Specifically, the formula strengthened host defense by modulating phagocytic activity in splenic macrophages, dendritic cells, and natural killer cells via Fcγ receptor-mediated pathways. Discussion: These findings provide mechanistic insights into artemisinin-associated immune potentiation. Moreover, we have proposed a novel ACT strategy targeting host-parasite interactions, offering a promising approach to circumvent emerging artemisinin resistance in Plasmodium species.