Multitarget Anticoccidial Activity of Olive Seed Phenolic Compounds Against Eimeria spp.: Insights from Molecular Docking and In Vitro Validation

Abstract

Purpose

The present investigation evaluates the anticoccidial potential of olive (Olea europaea L. var. Chemlal) seed ethanolic extract through in vitro assays and complementary in silico analyses against Eimeria spp.

Methods

The extract’s inhibitory effect was assessed on Eimeria spp. sporulated oocysts through microscopic and morphological evaluation of structural damage after 24 h of incubation, while molecular docking simulations were performed to predict the interactions of eight phenolic constituents identified by HPLC–ESI–MS with three Eimeria spp. target proteins including calcium-dependent protein kinase (PDB ID:4YSM), Extracellular Solute-Binding Protein Family 1 (PDB ID: 5IXP), and hexokinase (PDB ID: 6KSR).

Results

The in vitro results revealed that the explored extract significantly (p ≤ 0.05) disrupted the integrity of Eimeria oocysts at a concentration of 330 μg/mL, causing damage to their surface integrity and cell fragmentation after 24 h of incubation. Among the phenolics studied, four exhibited high binding affinities (-7.0 kcal/mol to -9.6 kcal/mol) for major parasitic targets. Three compounds demonstrated a notable multi-target profile. In this regard, 2,3 dihydro-amentoflavone interacted with the three proteins; isorhamnetin, oleuropein and 3-p-coumaryolquinic acid had affinity against two targets. This multitarget interaction pattern demonstrates olive seed chemicals' ability to affect multiple antiparasitic pathways simultaneously.

Conclusion

This integrated experimental and computational approach highlights the value of olive seed derivatives in developing alternative antiparasitic strategies.