title: Dihydropyrimidinone positive modulation of delta-subunit-containing gamma-aminobutyric acid type A receptors, including an epilepsy-linked mutant variant.
abstract: Gamma-aminobutyric acid type A receptors (GABA(A) receptors) are ligand-gated chloride channels that play a central role in signal transmission within the mammalian central nervous system. Compounds that modulate specific GABA(A) receptor subtypes containing the delta-subunit are scarce but would be valuable research tools and starting points for potential therapeutic agents. Here we report a class of dihydropyrimidinone (DHPM) heterocycles that preferentially potentiate peak currents of recombinant GABA(A) receptor subtypes containing the delta-subunit expressed in HEK293T cells. Using the three-component Biginelli reaction, 13 DHPMs with structural features similar to those of the barbiturate phenobarbital were synthesized; one DHPM used (monastrol) is commercially available. An up to approximately 3-fold increase in the current from recombinant alpha1beta2delta receptors was observed with the DHPM compound JM-II-43A or monastrol when co-applied with saturating GABA concentrations, similar to the current potentiation observed with the nonselective potentiating compounds phenobarbital and tracazolate. No agonist activity was observed for the DHPMs at the concentrations tested. A kinetic model was used in conjunction with dose-dependent measurements to calculate apparent dissociation constant values for JM-II-43A (400 muM) and monastrol (200 microM) at saturating GABA concentrations. We examined recombinant receptors composed of combinations of subunits alpha1, alpha4, alpha5, alpha6, beta2, beta3, gamma2L, and delta with JM-II-43A to demonstrate the preference for potentiation of delta-subunit-containing receptors. Lastly, reduced currents from receptors containing the mutated delta(E177A) subunit, described by Dibbens et al. [(2004) Hum. Mol. Genet. 13, 1315-1319] as a heritable susceptibility allele for generalized epilepsy with febrile seizures plus, are also potentiated by these DHPMs.
authors.affiliation: Department of Molecular Biology and Genetics, Biotechnology Building, Cornell University, Ithaca, New York 14853-2703, USA.
authors.forename: Ryan W
authors.initials: RW
authors.lastname: Lewis
authors.forename: John
authors.initials: J
authors.lastname: Mabry
authors.forename: Jason G
authors.initials: JG
authors.lastname: Polisar
authors.forename: Kyle P
authors.initials: KP
authors.lastname: Eagen
authors.forename: Bruce
authors.initials: B
authors.lastname: Ganem
authors.forename: George P
authors.initials: GP
authors.lastname: Hess
mesh_terms: D000409:Alanine / Q000235:genetics; D000494:Allosteric Regulation / Q000235:genetics; D002460:Cell Line; D004357:Drug Synergism; D004827:Epilepsy / Q000235:genetics* / Q000378:metabolism*; D014644:Genetic Variation; D018698:Glutamic Acid / Q000235:genetics; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding / Q000187:drug effects; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism*; D011743:Pyrimidines / Q000494:pharmacology; D011744:Pyrimidinones / Q000494:pharmacology*; D011963:Receptors, GABA-A / Q000235:genetics* / Q000378:metabolism*; D013871:Thiones / Q000494:pharmacology
chemical_list: C546240:GABRA1 protein, human; D020033:Protein Isoforms; D021122:Protein Subunits; D011743:Pyrimidines; D011744:Pyrimidinones; D011963:Receptors, GABA-A; D013871:Thiones; D018698:Glutamic Acid; C400223:monastrol; D000409:Alanine
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