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Updated: Jul 29, 2025
Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
Identification of novel anti-amoebic pharmacophores from kinase inhibitor chemotypes
Lori Ferrins1, Melissa J Buskes1, Madison M Kapteyn2
1Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, United States.
Abstract:
Acanthamoeba species, Naegleria fowleri, and Balamuthia mandrillaris are opportunistic pathogens that cause a range of brain, skin, eye, and disseminated diseases in humans and animals. These pathogenic free-living amoebae (pFLA) are commonly misdiagnosed and have sub-optimal treatment regimens which contribute to the extremely high mortality rates (>90%) when they infect the central nervous system. To address the unmet medical need for effective therapeutics, we screened kinase inhibitor chemotypes against three pFLA using phenotypic drug assays involving CellTiter-Glo 2.0. Herein, we report the activity of the compounds against the trophozoite stage of each of the three amoebae, ranging from nanomolar to low micromolar potency. The most potent compounds that were identified from this screening effort were: 2d (A. castellanii EC50: 0.92 ± 0.3 μM; and N. fowleri EC50: 0.43 ± 0.13 μM), 1c and 2b (N. fowleri EC50s: <0.63 μM, and 0.3 ± 0.21 μM), and 4b and 7b (B. mandrillaris EC50s: 1.0 ± 0.12 μM, and 1.4 ± 0.17 μM, respectively). With several of these pharmacophores already possessing blood-brain barrier (BBB) permeability properties, or are predicted to penetrate the BBB, these hits present novel starting points for optimization as future treatments for pFLA-caused diseases.
Insights
New kinase inhibitors show promise against deadly brain-infecting amoebae. This study screened compounds to find effective treatments for pathogenic free-living amoebae (pFLA) infections, offering hope for better patient outcomes.
Area of Science:
- Infectious Diseases
- Drug Discovery
- Parasitology
Background:
- Pathogenic free-living amoebae (pFLA) like *Acanthamoeba* species, *Naegleria fowleri*, and *Balamuthia mandrillaris* cause severe human and animal diseases.
- High mortality rates (>90%) for central nervous system infections are linked to misdiagnosis and inadequate treatments.
- There is a critical unmet need for effective therapeutics against pFLA.
Purpose of the Study:
- To screen kinase inhibitor chemotypes for activity against three pFLA species.
- To identify potent compounds for potential development into new treatments for pFLA infections.
- To address the limitations of current therapeutic options for these devastating diseases.
Main Methods:
- Phenotypic drug screening assays using CellTiter-Glo 2.0 were employed.
- Kinase inhibitor chemotypes were tested against the trophozoite stage of *Acanthamoeba* species, *Naegleria fowleri*, and *Balamuthia mandrillaris*.
- Compound potency was determined by measuring EC50 values.
Main Results:
- Several compounds demonstrated nanomolar to low micromolar potency against pFLA trophozoites.
- Compound 2d showed significant activity against *A. castellanii* (EC50: 0.92 ± 0.3 μM) and *N. fowleri* (EC50: 0.43 ± 0.13 μM).
- Compounds 1c, 2b, 4b, and 7b also exhibited potent activity against *N. fowleri* and *B. mandrillaris*.
Conclusions:
- Identified kinase inhibitors represent promising starting points for developing novel therapeutics against pFLA.
- Some compounds possess or are predicted to have blood-brain barrier permeability, crucial for treating CNS infections.
- Further optimization of these hits could lead to effective treatments for diseases caused by pFLA.
