PUBLICATION
A multiorganism pipeline for antiseizure drug discovery: Identification of chlorothymol as a novel γ-aminobutyric acidergic anticonvulsant
- Authors
- Jones, A., Barker-Haliski, M., Ilie, A.S., Herd, M.B., Baxendale, S., Holdsworth, C.J., Ashton, J.P., Placzek, M., Jayasekera, B.A.P., Cowie, C.J.A., Lambert, J.J., Trevelyan, A.J., White, H.S., Marson, A.G., Cunliffe, V.T., Sills, G.J., Morgan, A.
- ID
- ZDB-PUB-200817-14
- Date
- 2020
- Source
- Epilepsia 61(10): 2106-2118 (Journal)
- Registered Authors
- Baxendale, Sarah, Cunliffe, Vincent, Holdsworth, CJ
- Keywords
- GABA, drug discovery, epilepsy, nematode, zebrafish
- MeSH Terms
-
- Animals
- Anticonvulsants/chemistry*
- Anticonvulsants/pharmacology
- Anticonvulsants/therapeutic use*
- Caenorhabditis elegans
- Dose-Response Relationship, Drug
- Drug Discovery/methods*
- Drug Discovery/trends
- Female
- GABA-A Receptor Agonists/chemistry*
- GABA-A Receptor Agonists/pharmacology
- GABA-A Receptor Agonists/therapeutic use*
- Humans
- Male
- Mice
- Mice, Inbred C57BL
- Organ Culture Techniques
- Receptors, GABA-A/metabolism*
- Seizures/drug therapy*
- Seizures/genetics
- Seizures/metabolism
- Species Specificity
- Thymol/chemistry
- Thymol/pharmacology
- Thymol/therapeutic use
- Zebrafish
- PubMed
- 32797628 Full text @ Epilepsia
Citation
Jones, A., Barker-Haliski, M., Ilie, A.S., Herd, M.B., Baxendale, S., Holdsworth, C.J., Ashton, J.P., Placzek, M., Jayasekera, B.A.P., Cowie, C.J.A., Lambert, J.J., Trevelyan, A.J., White, H.S., Marson, A.G., Cunliffe, V.T., Sills, G.J., Morgan, A. (2020) A multiorganism pipeline for antiseizure drug discovery: Identification of chlorothymol as a novel γ-aminobutyric acidergic anticonvulsant. Epilepsia. 61(10):2106-2118.
Abstract
Objective Current medicines are ineffective in approximately one-third of people with epilepsy. Therefore, new antiseizure drugs are urgently needed to address this problem of pharmacoresistance. However, traditional rodent seizure and epilepsy models are poorly suited to high-throughput compound screening. Furthermore, testing in a single species increases the chance that therapeutic compounds act on molecular targets that may not be conserved in humans. To address these issues, we developed a pipeline approach using four different organisms.
Methods We sequentially employed compound library screening in the zebrafish, Danio rerio, chemical genetics in the worm, Caenorhabditis elegans, electrophysiological analysis in mouse and human brain slices, and preclinical validation in mouse seizure models to identify novel antiseizure drugs and their molecular mechanism of action.
Results Initially, a library of 1690 compounds was screened in an acute pentylenetetrazol seizure model using D rerio. From this screen, the compound chlorothymol was identified as an effective anticonvulsant not only in fish, but also in worms. A subsequent genetic screen in C elegans revealed the molecular target of chlorothymol to be LGC-37, a worm γ-aminobutyric acid type A (GABAA ) receptor subunit. This GABAergic effect was confirmed using in vitro brain slice preparations from both mice and humans, as chlorothymol was shown to enhance tonic and phasic inhibition and this action was reversed by the GABAA receptor antagonist, bicuculline. Finally, chlorothymol exhibited in vivo anticonvulsant efficacy in several mouse seizure assays, including the 6-Hz 44-mA model of pharmacoresistant seizures.
Significance These findings establish a multiorganism approach that can identify compounds with evolutionarily conserved molecular targets and translational potential, and so may be useful in drug discovery for epilepsy and possibly other conditions.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping