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Purpose <p>Malaria and leishmaniasis remain major global health challenges, exacerbated by increasing resistance to current antiparasitic drugs. Medicinal plants such as <i>Rhaponticum repens</i> and <i>Achillea millefolium</i> represent potential sources of new antiparasitic compounds, although their activity against <i>Plasmodium falciparum</i> and <i>Leishmania major</i> remains insufficiently validated. To evaluate the in vitro antiparasitic activity, cytotoxicity, and phytochemical composition of methanolic extracts of <i>R. repens</i> and <i>A. millefolium</i> against <i>P. falciparum</i> and <i>L. major </i>clinical isolates.</p> Methods <p>Methanolic extracts were tested in vitro against <i>P. falciparum</i> and <i>L. major</i>. Antiparasitic activity was assessed using JC-1 fluorescent dye staining followed by fluorescence quantification and flow cytometry, with chloroquine and amphotericin B as positive controls. Cytotoxicity was evaluated on Vero cells using the MTT assay. Phytochemical profiling was performed by GC–MS. IC₅₀ values and selectivity indices were calculated from triplicate experiments.</p> Results <p>Both extracts exhibited significant dose- and time-dependent inhibition of parasite viability. <i>R. repens</i> showed higher activity against <i>L. major</i> (81% inhibition at 2.0 mg/mL, IC₅₀ = 1.18 mg/mL; SI = 10.6), whereas <i>A. millefolium</i> was more effective against <i>P. falciparum</i> (94% inhibition at 2.0 mg/mL, IC₅₀ = 0.52 mg/mL; SI = 26.3). Cytotoxicity toward mammalian cells was low. GC–MS analysis identified nonadecane and docosane as major constituents.</p> Conclusion <p>Methanolic extracts of <i>R. repens</i> and <i>A. millefolium</i> demonstrate promising in vitro antiparasitic activity with acceptable safety profiles. These findings support their potential as leads for natural product-based antiparasitic drug discovery and provide a basis for future bio-guided fractionation studies.</p>

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In Vitro Antiparasitic Activity and Phytochemical Profiling of Rhaponticum repens and Achillea millefolium Methanolic Extracts Against Plasmodium Falciparum and Leishmania major

  • 期刊论文

  • 发表日期：

    2026年2月16日

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  • Kiran Singh,
  • Abolghasem Siyadatpanah,
  • Abdulaziz Garba,
  • Jitendra Singh,
  • Roghayeh Norouzi,
  • Mourad Ben Said,
  • Hanène Belkahia

摘要

Purpose

Malaria and leishmaniasis remain major global health challenges, exacerbated by increasing resistance to current antiparasitic drugs. Medicinal plants such as Rhaponticum repens and Achillea millefolium represent potential sources of new antiparasitic compounds, although their activity against Plasmodium falciparum and Leishmania major remains insufficiently validated. To evaluate the in vitro antiparasitic activity, cytotoxicity, and phytochemical composition of methanolic extracts of R. repens and A. millefolium against P. falciparum and L. major clinical isolates.

Methods

Methanolic extracts were tested in vitro against P. falciparum and L. major. Antiparasitic activity was assessed using JC-1 fluorescent dye staining followed by fluorescence quantification and flow cytometry, with chloroquine and amphotericin B as positive controls. Cytotoxicity was evaluated on Vero cells using the MTT assay. Phytochemical profiling was performed by GC–MS. IC₅₀ values and selectivity indices were calculated from triplicate experiments.

Results

Both extracts exhibited significant dose- and time-dependent inhibition of parasite viability. R. repens showed higher activity against L. major (81% inhibition at 2.0 mg/mL, IC₅₀ = 1.18 mg/mL; SI = 10.6), whereas A. millefolium was more effective against P. falciparum (94% inhibition at 2.0 mg/mL, IC₅₀ = 0.52 mg/mL; SI = 26.3). Cytotoxicity toward mammalian cells was low. GC–MS analysis identified nonadecane and docosane as major constituents.

Conclusion

Methanolic extracts of R. repens and A. millefolium demonstrate promising in vitro antiparasitic activity with acceptable safety profiles. These findings support their potential as leads for natural product-based antiparasitic drug discovery and provide a basis for future bio-guided fractionation studies.