Query String: FLUMAZENIL
BDBM26264 FLUMAZENIL
BDBM26263 Romazicon CHEMBL407 Anexate Ro15-1788 FLUMAZENIL [3H]Ro15-1788 ethyl 12-fluoro-8-methyl-9-oxo-2,4,8-triazatricyclo[8.4.0.0^{2,6}]tetradeca-1(14),3,5,10,12-pentaene-5-carboxylate
- ChEMBL_2280319 Displacement of flumazenil from alpha5 GABAA receptor (unknown origin)
- ChEMBL_2280320 Displacement of flumazenil from alpha1 GABAA receptor (unknown origin)
- ChEMBL_1901505 (CHEMBL4403727) Displacement of [3H]-flumazenil from human GABAA alpha1beta3gamma2 expressed in Ltk cells
- ChEMBL_1901507 (CHEMBL4403729) Displacement of [3H]-flumazenil from human GABAA alpha3beta3gamma2 expressed in Ltk cells
- ChEMBL_1901508 (CHEMBL4403730) Displacement of [3H]-flumazenil from human GABAA alpha5beta3gamma2 expressed in Ltk cells
- ChEMBL_69640 (CHEMBL676828) Inhibition of [3H]-Flumazenil binding to recombinant GABA-A receptor alpha-1 subunit in cerebellum
- ChEMBL_69806 (CHEMBL677202) Inhibition of [3H]flumazenil binding to recombinant GABA-A receptor alpha-5 subunit in cerebellum
- ChEMBL_861270 (CHEMBL2173576) Displacement of [3H]flumazenil from human recombinant GABA-A alpha5beta3gamma2L receptor in HEK293 cells
- ChEMBL_623168 (CHEMBL1111448) Displacement of [3H]flumazenil from GABAA alpha-5-beta-3-gamma-2 receptor
- ChEMBL_623169 (CHEMBL1111449) Displacement of [3H]flumazenil from GABAA alpha-1-beta-3-gamma-2 receptor
- ChEMBL_623170 (CHEMBL1111450) Displacement of [3H]flumazenil from GABAA alpha-2-beta-3-gamma-2 receptor
- ChEMBL_623171 (CHEMBL1111451) Displacement of [3H]flumazenil from GABAA alpha-3-beta-3-gamma-2 receptor
- ChEMBL_69641 (CHEMBL676829) Inhibition of [3H]flumazenil binding to recombinant GABA-A receptor alpha-1 subunit in spinal cord
- ChEMBL_69807 (CHEMBL677203) Inhibition of [3H]-Flumazenil binding to recombinant GABA-A receptor alpha-5 subunit in spinal cord
- ChEMBL_726365 (CHEMBL1687497) Displacement of [3H]flumazenil from GABAA receptor subunit alpha-1 expressed in CHO cells
- ChEMBL_726366 (CHEMBL1687498) Displacement of [3H]flumazenil from GABAA receptor subunit alpha-2 expressed in CHO cells
- ChEMBL_861269 (CHEMBL2173575) Displacement of [3H]flumazenil from human recombinant GABA-A alpha2beta2gamma2L receptor expressed in HEK293 cells
- ChEMBL_1747527 (CHEMBL4182037) Displacement of [3H]-flumazenil from human GABA-A receptor alpha1beta2gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747528 (CHEMBL4182038) Displacement of [3H]-flumazenil from rat GABA-A receptor alpha1beta2gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747529 (CHEMBL4182039) Displacement of [3H]-flumazenil from human GABA-A receptor alpha2beta3gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747530 (CHEMBL4182040) Displacement of [3H]-flumazenil from rat GABA-A receptor alpha2beta3gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747531 (CHEMBL4182041) Displacement of [3H]-flumazenil from human GABA-A receptor alpha3beta3gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747532 (CHEMBL4182042) Displacement of [3H]-flumazenil from rat GABA-A receptor alpha3beta3gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747533 (CHEMBL4182043) Displacement of [3H]-flumazenil from human GABA-A receptor alpha5beta3gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_1747534 (CHEMBL4182044) Displacement of [3H]-flumazenil from rat GABA-A receptor alpha5beta3gamma2 expressed in HEK293 cells by electrophysiology assay
- ChEMBL_2275518 Displacement of [3H]-flumazenil from rat recombinant alpha1 GABAA receptor assessed as inhibition constant in presence of zolpidem
- ChEMBL_2275519 Displacement of [3H]-flumazenil from rat recombinant alpha2 GABAA receptor assessed as inhibition constant in presence of zolpidem
- ChEMBL_69958 (CHEMBL678631) Inhibition of [3H]flumazenil binding to recombinant GABA-A receptor alpha-1-beta-2-gamma-2 subunits
- ChEMBL_343682 (CHEMBL861092) Displacement of [3H]flumazenil from rat GABA-Aalpha1 receptor plus beta-2-gamma-2 in HEK293 cells
- ChEMBL_343683 (CHEMBL861093) Displacement of [3H]flumazenil from rat GABA-Aalpha2 receptor plus beta-2-gamma-2 in HEK293 cells
- ChEMBL_343684 (CHEMBL861094) Displacement of [3H]flumazenil from rat GABA-Aalpha5 receptor plus beta-3-gamma-2 in HEK293 cells
- ChEMBL_428867 (CHEMBL917143) Displacement of [3H]flumazenil from rat GABAA alpha-1-beta-2-gamma-2 expressed in HEK293 cells
- ChEMBL_428868 (CHEMBL917144) Displacement of [3H]flumazenil from rat GABAA alpha-2-beta-2-gamma-2 expressed in HEK293 cells
- ChEMBL_428870 (CHEMBL917146) Displacement of [3H]flumazenil from rat GABAA alpha-5-beta-3-gamma-2 expressed in HEK293 cells
- ChEMBL_492498 (CHEMBL950411) Displacement of [3H]flumazenil in rat GABAA alpha-1-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_492499 (CHEMBL950412) Displacement of [3H]flumazenil in rat GABAA alpha-2-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_492500 (CHEMBL950413) Displacement of [3H]flumazenil in rat GABAA alpha-3-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_492501 (CHEMBL951364) Displacement of [3H]flumazenil in rat GABAA alpha-5-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_341422 (CHEMBL861815) Displacement of [3H]Flumazenil from human GABA-Aalpha1 receptor plus beta-2-gamma-2 expressed in HEK293 cells
- ChEMBL_341423 (CHEMBL861816) Displacement of [3H]Flumazenil from human GABA-Aalpha3 receptor plus beta-2-gamma-2 expressed in HEK293 cells
- ChEMBL_519946 (CHEMBL954625) Displacement of [3H]flumazenil from rat GABA-A alpha-1-beta-2-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_519947 (CHEMBL954626) Displacement of [3H]flumazenil from rat GABA-A alpha-2-beta-2-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_519949 (CHEMBL954628) Displacement of [3H]flumazenil from rat GABA-A alpha-5-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_217124 (CHEMBL822944) Inhibition of [3H]flumazenil binding to rat alpha-1-beta-2-gamma-2 GABA-A receptor subunits expressed in HEK293 cells
- ChEMBL_217125 (CHEMBL822945) Inhibition of [3H]flumazenil binding to rat alpha-1-beta-2-gamma-2 GABA-A receptor subunits expressed in HEK293 cells
- ChEMBL_217291 (CHEMBL823759) Inhibition of [3H]flumazenil binding to rat alpha-3-beta-2-gamma-2 GABA-A receptor subunits expressed in HEK293 cells
- ChEMBL_217292 (CHEMBL823760) Inhibition of [3H]flumazenil binding to rat alpha-3-beta-2-gamma-2 GABA-A receptor subunits expressed in HEK293 cells
- ChEMBL_217645 (CHEMBL818517) Inhibition of [3H]flumazenil binding to rat alpha-5-beta-3-gamma-2 GABA-A receptor subunits expressed in HEK293 cells
- ChEMBL_217646 (CHEMBL818518) Inhibition of [3H]flumazenil binding to rat alpha-5-beta-3-gamma-2 GABA-A receptor subunits expressed in HEK293 cells
- ChEMBL_491163 (CHEMBL992851) Displacement of [3H]flumazenil from human recombinant GABAA alpha-1-beta-2-gamma-2S receptor expressed in HEK293 cells
- ChEMBL_619498 (CHEMBL1111355) Displacement of [3H]Flumazenil from cloned human GABA alpha-5-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_619504 (CHEMBL1111361) Displacement of [3H]Flumazenil from human GABA alpha-5-beta-3-gamma-2 receptor expressed in insect SF9 cells
- ChEMBL_619508 (CHEMBL1111365) Displacement of [3H]Flumazenil from cloned human GABA alpha-1-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_619509 (CHEMBL1111366) Displacement of [3H]Flumazenil from cloned human GABA alpha-2-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_619510 (CHEMBL1111367) Displacement of [3H]Flumazenil from cloned human GABA alpha-3-beta-3-gamma-2 receptor expressed in HEK293 cells
- ChEMBL_70394 (CHEMBL683020) Inhibition of [3H]-flumazenil binding to rat GABA-A receptor alpha-3-beta-2-gamma-2 subunits expressed in HEK293 cells
- ChEMBL_69455 (CHEMBL682784) Inhibition of [3H]flumazenil specific binding at rat GABA-A alpha-1-beta-2-gamma-2 receptor subunits expressed in HEK293 cells
- ChEMBL_701327 (CHEMBL1648947) Displacement of [3H]flumazenil human recombinant alpha3beta3gamma2 GABA(A) receptor expressed in HEK293 cells after 30 mins by glass fiber filtration assay
- ChEMBL_701328 (CHEMBL1648948) Displacement of [3H]flumazenil human recombinant alpha2beta3gamma2 GABA(A) receptor expressed in HEK293 cells after 30 mins by glass fiber filtration assay
- ChEMBL_701329 (CHEMBL1648949) Displacement of [3H]flumazenil human recombinant alpha1beta3gamma2 GABA(A) receptor expressed in HEK293 cells after 30 mins by glass fiber filtration assay
- ChEMBL_701331 (CHEMBL1648951) Displacement of [3H]flumazenil human recombinant alpha5beta3gamma2 GABA(A) receptor expressed in HEK293 cells after 30 mins by glass fiber filtration assay
- ChEMBL_1901493 (CHEMBL4403715) Displacement of [3H]-flumazenil from human GABAA alpha1beta3gamma2 expressed in HEK293 cell membranes measured after 2 hrs by liquid scintillation counting method
- ChEMBL_1901499 (CHEMBL4403721) Displacement of [3H]-flumazenil from human GABAA alpha3beta3gamma2 expressed in HEK293 cell membranes measured after 2 hrs by liquid scintillation counting method
- ChEMBL_1901500 (CHEMBL4403722) Displacement of [3H]-flumazenil from human GABAA alpha5beta3gamma2 expressed in HEK293 cell membranes measured after 2 hrs by liquid scintillation counting method
- ChEMBL_303742 (CHEMBL829785) Displacement of [3H]flumazenil from rat GABA-A Gamma-aminobutyric acid A receptor alpha-1-beta-2-gamma-2 expressed in HEK293 cells
- ChEMBL_303743 (CHEMBL829786) Displacement of [3H]flumazenil from rat GABA-A Gamma-aminobutyric acid A receptor alpha-2-beta-2-gamma-2 expressed in HEK293 cells
- ChEMBL_303744 (CHEMBL829787) Displacement of [3H]flumazenil from rat GABA-A Gamma-aminobutyric acid A receptor alpha-5-beta-3-gamma-2 expressed in HEK293 cells
- ChEMBL_1850878 (CHEMBL4351502) Displacement of [3H]-flumazenil from human GABAA alpha3beta3gamma2 receptor expressed in HEK293 cell membranes measured after 2 hrs by liquid scintillation counting method
- ChEMBL_1850883 (CHEMBL4351507) Displacement of [3H]-flumazenil from human GABAA alpha1beta3gamma2 receptor expressed in HEK293 cell membranes measured after 2 hrs by liquid scintillation counting method
- ChEMBL_1850887 (CHEMBL4351511) Displacement of [3H]-flumazenil from human GABAA alpha5beta2gamma2 receptor expressed in HEK293 cell membranes measured after 2 hrs by liquid scintillation counting method
- Binding Assay for gamma2-containing GABAA Subtypes The affinity of compounds at GABAA γ2 subunit-containing receptors was measured by competition for [3H]Flumazenil (81.1 Ci/mmol; Roche) binding to HEK293F cells expressing human (transiently transfected) receptors of composition α1β3γ2.
- Binding Assay The affinity of compounds at GABAA receptor subtypes was measured by competition for [3H]flumazenil (85 Ci/mmol; Roche) binding to HEK293 cells expressing rat (stably transfected) or human (transiently transfected) receptors of composition α1β3γ2, α2β3γ2, α3β3γ2 and α5β3γ2.
- Binding Assay The affinity of the test compounds was determined by radioligand competition binding assay, using the known compound [3H]Ro-15-1788 (Flumazenil) (Perkin Elmer, 85.4 Ci/mmol) and the human recombinant GABA A receptor containing the alpha2, beta2, and gamma2 subunits.Membranes were prepared from HEK cells expressing hGABA A alpha2beta2-gamma2 receptor, and validated to ascertain protein concentration, receptor expression and to determine the Kd of the flumazenil as well as the Ki of a standard set of compounds before being used to test new compounds.The assay was carried out in 96 well plates; testing compounds using a 10 point semi-log dilution range from 19 uM top concentration. 100 ul of radioligand and 100 ul of membrane in 50 mM Tris-HCl and 0.05% F127 with 1 ul of test compound was incubated for 2 hours to allow the reaction to achieve equilibrium, and then harvested onto filter plates, dried and counted on a TopCount NXT. The data was analysed, and the Ki values were presented as the geometric mean of at least two replicates.
- Binding Assay The affinity of the test compounds was determined by radioligand competition binding assay, using the known compound [3H]Ro-15-1788 (Flumazenil) (Perkin Elmer, 85.4 Ci/mmol) and the human recombinant GABA A receptor containing the alpha2, beta2, and gamma3 subunits.Membranes were prepared from HEK cells expressing hGABA A alpha2beta2-gamma3 receptor, and validated to ascertain protein concentration, receptor expression and to determine the Kd of the flumazenil as well as the Ki of a standard set of compounds before being used to test new compounds.The assay was carried out in 96 well plates; testing compounds using a 10 point semi-log dilution range from 19 uM top concentration. 100 ul of radioligand and 100 ul of membrane in 50 mM Tris-HCl and 0.05% F127 with 1 ul of test compound was incubated for 2 hours to allow the reaction to achieve equilibrium, and then harvested onto filter plates, dried and counted on a TopCount NXT. The data was analysed, and the Ki values were presented as the geometric mean of at least two replicates.
- Radioligand Competition Binding Assay The affinity of the test compounds was determined by radioligand competition binding assay, using the known compound [3H]Ro-15-1788 (Flumazenil) (Perkin Elmer, 85.4 Ci/mmol) and the human recombinant GABA A receptor containing the alpha2, beta2, and gamma3 subunits.Membranes were prepared from HEK cells expressing hGABA A alpha2beta2- gamma3 receptor, and validated to ascertain protein concentration, receptor expression and to determine the Kd of the flumazenil as well as the Ki of a standard set of compounds before being used to test new compounds.The assay was carried out in 96 well plates; testing compounds using a 10 point semi-log dilution range from 19 uM top concentration. 100 ul of radioligand and 100 ul of membrane in 50 mM Tris-HCI and 0.05% F127 with 1 ul of test compound was incubated for 2 hours to allow the reaction to achieve equilibrium, and then harvested onto filter plates, dried and counted on a TopCount NXT. The data was analysed, and the Ki values were presented.
- Binding Assay for γ2-Containing GABAA Subtypes Table 1: Radioligand binding assays were carried out in a volume of 200 μL (96-well plates) which contained 100 μL of cell membranes, [3H]Flumazenil at a concentration of 1 nM and the test compound in the range of 0.1×10−9 to 30×10−6 M. Nonspecific binding was defined by 10−5 M Diazepam and typically represented less than 5% of the total binding. Assays were incubated to equilibrium for 1 hour at 4° C. and harvested onto GF/C uni-filters (Packard) by filtration using a Packard harvester and washing with ice-cold wash buffer (50 mM Tris; pH 7.5). After drying, filter-retained radioactivity was detected by liquid scintillation counting. Ki values were calculated using Exel-Fit (Microsoft) and are the means of two determinations. The compounds of the accompanying examples were tested in the above described assay, and the preferred compounds were found to possess large K, value for displacement of [3H]Flumazenil from the α1β3γ2 subtype of the human GABAA receptor of 100 nM or above. Most preferred are compounds with a Ki α1β3γ2 (nM)>300.
- Binding Assay for γ2-Containing GABAA Subtypes Table 1: Radioligand binding assays were carried out in a volume of 200 uL (96-well plates) which contained 100 uL of cell membranes, [3H]Flumazenil at a concentration of 1 nM and the test compound in the range of [0.1 10+-3+-10]??10+-6 M. Nonspecific binding was defined by 10+-5 M Diazepam and typically represented less than 5% of the total binding. Assays were incubated to equilibrium for 1 hour at 4?? C. and harvested onto GF/C uni-filters (Packard) by filtration using a Packard harvester and washing with ice-cold wash buffer (50 mM Tris; pH 7.5). After anhydrousing, filter-retained radioactivity was detected by liquid scintillation counting. Ki values were calculated using Excel-Fit (Microsoft) and are the means of two determinations.The compounds of the accompanying examples were tested in the above described assay, and the preferred compounds were found to possess large Ki value for displacement of [3H]Flumazenil from the alpha1beta3gamma 2 subtype of the human GABAA receptor of 100 nM or above. Most preferred are compounds with a Ki alpha1beta3gamma 2 (nM) >300. In a preferred embodiment the compounds of the invention are binding selectively for the gamma 1 subunit-containing GABAA receptors relative to gamma 2 subunit-containing GABAA receptors.
- Radioligand Binding Assay Radioligand binding assays were carried out in a volume of 200 mL (96-well plates) which contained 100 mL of cell memebranes, [3H]flumazenil at a concentration of 1 nM for α1, α2, α3 subunits and 0.5 nM for α5 subunits and the test compound in the range of 10-10−3x10−6 M. Nonspecific binding was defined by 10−5 M diazepam and typically represented less than 5% of the total binding. Assays were incubated to equilibrium for 1 hour at 4 C. and harvested onto GF/C uni-filters (Packard) by filtration using a Packard harvester and washing with ice-cold wash buffer (50 mM Tris; pH 7.5). After drying, filter-retained radioactivity was detected by liquid scintillation counting.
- Radioligand Binding Assay Radioligand binding assays were carried out in a volume of 200 mL (96-well plates) which contained 100 mL of cell membranes, [3H]flumazenil at a concentration of 1 nM for α1, α2, α3 subunits and 0.5 nM for α5 subunits and the test compound in the range of 10-10−3×10−6 M. Nonspecific binding was defined by 10−5 M diazepam and typically represented less than 5% of the total binding. Assays were incubated to equilibrium for 1 hour at 4° C. and harvested onto GF/C uni-filters (Packard) by filtration using a Packard harvester and washing with ice-cold wash buffer (50 mM Tris; pH 7.5). After drying, filter-retained radioactivity was detected by liquid scintillation counting.