Molecular and Pharmacological Evaluation of a Curcuma longa, Zingiber officinale and Allium sativum Polyherbal Formulation: Unravelling Synergistic Mechanisms against Plasmodium berghei

Ethnopharmacological relevance: Curcuma longa (turmeric), Zingiber officinale (ginger), and Allium sativum (garlic) have a long history of use in traditional medicine, frequently formulated together as decoctions or herbal teas to manage febrile illnesses and malaria. However, despite widespread ethnomedicinal use, scientific evidence on their combined pharmacological efficacy and mechanisms remains scarce.. Aim of the study: This study aims to elucidate the synergistic molecular and pharmacological mechanisms of a Curcuma longa , Zingiber officinale , and Allium sativum polyherbal formulation against Plasmodium berghei . Materials and methods: The ethanol-extracted polyherbal blend was analysed by GC-MS, revealing 21 bioactive compounds—oleic acid (15.84%), squalene (8.43%), ar-turmerone (6.24%), and Cholest-14-en-3-ol (4.76%) as major constituents. Molecular docking against Plasmodium falciparum chloroquine resistance transporter (PfCRT, PDB ID: 6UKJ) showed strong binding affinities for Cholest-14-en-3-ol and dehydroabietol (–8.4 kcal/mol), outperforming chloroquine (–5.4 kcal/mol). Pharmacophore modelling revealed critical hydrophobic and aromatic interactions. Stability of protein-ligand complexes was validated by 100 ns molecular dynamics simulations and MM/PBSA analysis (ΔG_bind = –33.13 kcal/mol). Acute oral toxicity was assessed in mice at doses up to 5000 mg/kg. Results: The extract significantly reduced Plasmodium berghei -induced parasitemia by 97% on day 4, comparable to chloroquine (98%) (p<0.05). Haematological and liver function parameters normalised, while antioxidant markers (SOD, GPx) improved and lipid peroxidation (MDA) decreased. Conclusion: This polyherbal formulation exhibits potent antiplasmodial and antioxidant effects, likely mediated via synergistic interactions of its phytoconstituents with parasitic molecular targets. It presents a promising ethnomedicine-inspired candidate for alternative malaria therapy, especially in drug-resistant contexts.