Development and optimization of a high-throughput screening method utilizing Ancylostoma ceylanicum egg hatching to...

Area of Science:

• * Parasitology and Tropical Medicine
• * Drug Discovery and Development

Background:

• * Hookworm infections are a significant global health issue, affecting hundreds of millions and necessitating new chemotherapies.
• * Developing novel anthelmintics requires identifying essential parasite targets and effective drug candidates through rigorous testing.
• * Conventional drug discovery methods are often slow and expensive, highlighting the need for efficient screening techniques.

Purpose of the Study:

• * To establish a high-throughput screening (HTS) method for identifying novel anthelmintic compounds against hookworms.
• * To screen a large library of small molecules for their ability to inhibit the egg hatching of *Ancylostoma ceylanicum*.
• * To evaluate the safety and efficacy of identified compounds by assessing their toxicity to human cells and determining their inhibitory concentrations.

Main Methods:

• * A novel high-throughput, plate reader-based assay was developed to assess compound toxicity against *Ancylostoma ceylanicum* L1 larvae.
• * 39,568 compounds were screened at 10 μM, with subsequent verification of nematicidal activity and human cell line (HeLa) toxicity for hit compounds.
• * Half-maximal inhibitory concentration (IC50) values were determined for the most promising compounds.

Main Results:

• * The HTS identified 830 compounds inhibiting *A. ceylanicum* egg hatching by >50%, with 132 compounds showing >90% inhibition.
• * Fifty-nine compounds demonstrated significant hookworm egg hatching inhibition (>80%) with minimal toxicity (<20%) to human HeLa cells.
• * IC50 values for these 59 compounds ranged from 0.05-8.94 μM, indicating potent activity.

Conclusions:

• * The developed HTS assay is effective for rapidly identifying potential anthelmintic drug candidates.
• * The study identified 59 promising compounds with validated nematicidal activity and favorable safety profiles for further drug development.
• * These findings contribute valuable information for the development of new treatments for human hookworm disease.