title: The proconvulsant effects of the GABAA alpha5 subtype-selective compound RY-080 may not be alpha5-mediated.
abstract: RY-080 (ethyl 8-ethynyl-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine-3-carboxylate) is an imidazobenzodiazepine with 40-50-fold higher affinity for the benzodiazepine binding site of alpha5- rather than alpha1-, alpha2- or alpha3-containing GABAA receptors. Previous data describing RY-080 as being convulsant suggests that inverse agonists selective for the alpha5 subtype may not be suitable for clinical development. In the present study, we show that RY-080 possesses inverse agonism for the alpha1 and alpha5 subtypes of human recombinant GABAA receptors and whilst not convulsant it was proconvulsant. Hence, with pentylenetetrazole alone, the dose predicted to give tonic convulsions in 50% of the mice (ED50) was 108 mg/kg whereas in the presence of 1 and 10 mg/kg RY-080, the ED50s were 93 and 57 mg/kg, respectively. In vivo [3H]L-655,708 and [3H]Ro 15-1788 binding assays showed that the subtype selectivity of RY-080 in vivo was 7-10-fold for alpha5-relative to alpha1- and alpha2/alpha3-containing receptors (respective ID50 values of 0.93, 9.7 and 6.2 mg/kg) and is therefore much lower than seen in vitro. Consequently, it is not possible to define a dose of RY-080 which gives high occupancy of the alpha5 subtype without binding to other subtypes and accordingly the proconvulsant effects of RY-080 cannot be attributed solely to the alpha5 subtype.
authors.affiliation: Merck Sharp and Dohme Research Laboratories, Neuroscience Research Centre, Terlings Park, Eastwick Road, Harlow, Essex CM20 2QR, England. John_Atack@Merck.com
authors.forename: John R
authors.initials: JR
authors.lastname: Atack
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Bayley
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Fletcher
authors.forename: Ruth M
authors.initials: RM
authors.lastname: McKernan
authors.forename: Keith A
authors.initials: KA
authors.lastname: Wafford
authors.forename: Gerard R
authors.initials: GR
authors.lastname: Dawson
mesh_terms: D000480:Alkynes / Q000494:pharmacology*; D000818:Animals; D001569:Benzodiazepines / Q000494:pharmacology*; D002478:Cells, Cultured; D003292:Convulsants / Q000494:pharmacology*; D005260:Female; D005442:Flumazenil / Q000378:metabolism; D018757:GABA Modulators / Q000378:metabolism; D058785:GABA-A Receptor Agonists; D007093:Imidazoles / Q000378:metabolism / Q000494:pharmacology*; D008024:Ligands; D008297:Male; D051379:Mice; D009865:Oocytes / Q000378:metabolism; D010433:Pentylenetetrazole / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000378:metabolism*; D012640:Seizures / Q000139:chemically induced / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D000480:Alkynes; D003292:Convulsants; D018757:GABA Modulators; D058785:GABA-A Receptor Agonists; C496250:Gabra5 protein, mouse; D007093:Imidazoles; C105490:L 655,708; D008024:Ligands; C109021:RY 80; D011963:Receptors, GABA-A; D001569:Benzodiazepines; D005442:Flumazenil; D010433:Pentylenetetrazole