Ursolic acid acetate and iso-mukaadial acetate bind to Plasmodium falciparum Hsp90, abrogating its chaperone function in vitro

• Published in: Naunyn-Schmiedeberg's archives of pharmacology 2024-07, Vol.397 (7), p.5179-5192
• Main Authors: Nndwammbi, Andani A. T, Dongola, Tendamudzimu Harmfree, Shonhai, Addmore, Mokoena, Fortunate, Pooe, Ofentse J., Simelane, Mthokozisi B. C
• Format: Article
• Language: English
• Subjects: Acetates - chemistry; Acetates - pharmacology; Acetic acid; Antimalarial activity; Antimalarial agents; Antimalarials - pharmacology; Antiparasitic agents; Antiprotozoal agents; Biomedical and Life Sciences; Biomedicine; Citrate synthase; Drug resistance; Fourier transforms; Heat shock proteins; HSP90 Heat-Shock Proteins - metabolism; Hsp90 protein; Malaria; Malate dehydrogenase; Molecular Docking Simulation; Molecular dynamics; Molecular Dynamics Simulation; Neurosciences; Pharmacology/Toxicology; Plasmodium falciparum; Plasmodium falciparum - drug effects; Protein Binding; Protein folding; Protein structure; Protozoan Proteins - metabolism; Secondary structure; Surface Plasmon Resonance; Therapeutic targets; Triterpenes - chemistry; Triterpenes - pharmacology; Ursolic Acid
• ISSN: 0028-1298; 1432-1912; 1432-1912
• DOI: 10.1007/s00210-024-02944-9

Plasmodium falciparum is the most lethal malaria parasite. Increasing incidences of drug resistance of P. falciparum have prompted the need for discovering new and effective antimalarial compounds with an alternative mode of action. Heat shock protein 90 (PfHsp90) facilitates protein folding and is a promising antimalarial drug target. We have previously reported that iso-mukaadial acetate (IMA) and ursolic acid acetate (UAA) exhibit antimalarial activity. We investigated the abilities of IMA and UAA to bind PfHsp90 by molecular docking and dynamics simulations. The in silico predictions were validated by biochemical assays conducted on recombinant PfHsp90. The interaction between the ligands and PfHsp90 was evaluated using ultraviolet-visible spectroscopy (UV-vis), Fourier transform infrared (FTIR), and surface plasmon resonance (SPR) analysis. The results obtained by docking calculations and MD dynamics simulation predicted that UAA and IMA preferentially bound to PfHsp90 via the N-terminal domain, with UAA binding more stable than IMA. UV-vis-based data suggest that PfHsp90 harbors buried aromatic amino acids, which were exposed in the presence of either IMA or UAA. In addition, data obtained using FTIR suggested that IMA and UAA destabilized the secondary structure of PfHsp90. Of the two compounds, UAA bound to PfHsp90 within the micromolar range based on surface plasmon resonance (SPR)-based binding assay. Furthermore, both compounds disrupted the holdase chaperone function of PfHsp90 as the chaperone failed to suppress heat-induced aggregation of the model proteins, malate dehydrogenase (MDH), luciferase, and citrate synthase in vitro . In addition, both compounds lowered the ATPase activity of PfHsp90. The molecular dynamics simulation analysis indicated that the docked complexes were mostly stable for 100 ns, validating the data obtained through the biochemical assays. Altogether, this study expands the repository of antiplasmodial compounds that have PfHsp90 among their possible targets.