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A FLIPR Assay for Discovery of GABAA Receptor Modulators of Natural Origin
- Source :
- Planta medica. 85(11-12)
- Publication Year :
- 2019
-
Abstract
- A fluorometric imaging plate reader (FLIPR) assay utilizing Chinese hamster ovary (CHO) cells stably transfected with GABAA receptors of α 1 β 2 γ 2 subunit composition was evaluated and validated for rapid screening of plant extract libraries and efficient localization of active compounds in extracts. Validation was performed with pure compounds and extracts known to contain allosteric GABAA receptor modulators. Plants extracts that had been previously reported as active in an assay using Xenopus laevis oocytes transiently expressing GABAA receptors of α 1 β 2 γ 2 subunit composition were also active in the FLIPR assay. A protocol for HPLC-based activity profiling was developed, whereby separations of 0.4 – 1.2 mg of extracts on an analytical HPLC column were found to be sufficient for the sensitivity of the bioassay. The protocol successfully localized the activity of known GABAergic natural products, such as magnolol in Magnolia officinalis, valerenic acid in Valeriana officinalis, and piperine in Piper nigrum extract. EC50 values of compounds (magnolol: 4.81 ± 1.0 µM, valerenic acid: 12.56 ± 1.2 µM, and piperine: 5.76 ± 0.7 µM) were found to be comparable or lower than those reported using Xenopus oocyte assays.
- Subjects :
- Polyunsaturated Alkamides
Allosteric regulation
Pharmaceutical Science
CHO Cells
Lignans
Analytical Chemistry
chemistry.chemical_compound
Xenopus laevis
Alkaloids
Cricetulus
Piperidines
Valerian
Drug Discovery
Animals
Fluorometry
Benzodioxoles
Chromatography, High Pressure Liquid
Pharmacology
biology
GABAA receptor
Plant Extracts
Chinese hamster ovary cell
Organic Chemistry
Biphenyl Compounds
Valerenic acid
biology.organism_classification
Receptors, GABA-A
Magnolol
Magnolia officinalis
Complementary and alternative medicine
chemistry
Biochemistry
Indenes
Magnolia
Piperine
Oocytes
Molecular Medicine
Biological Assay
Piper nigrum
Sesquiterpenes
Plate reader
Subjects
Details
- ISSN :
- 14390221
- Volume :
- 85
- Issue :
- 11-12
- Database :
- OpenAIRE
- Journal :
- Planta medica
- Accession number :
- edsair.doi.dedup.....7625b113c7734c30e3f316a30b5cd5d1