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Assay Detail

CHEMBL5733771

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Binding
Inhibition Assay: Cytochrome P450 3A4 and 2D6:Recombinant enzymes, 3A4 and 2D6, generated using the ABL yeast expression system were used. For CYP3A4, the enzyme amount was 2 pmol per well, and the substrate (7-benzyloxy-4-(trifluoromethyl) coumarin; BFC) was used at 15 μM. The assay mixture also included K2HPO4 (pH 7.4; conc. 0.1 M) and NADPH (conc. 1 mM). The incubation time was 30 minutes, and the reference inhibitor was A-naphthoflavone. For CYP2D6, the enzyme amount was 2 pmol per well, and the substrate (7-methoxy-4-(aminomethyl)-coumarin; MAMC) was used at 20 μM. The assay mixture also included K2HPO4 (pH 7.4; conc. 0.1 M) and NADPH (conc. 0.06 mM). The incubation time was 35 minutes, and the reference inhibitor was quinidine.Briefly, 0.6 μL of serially diluted compound was added to 75 μL water. 10 μL of diluted compound in water was then added to each assay plate. 20 μL of a K2HPO4, enzyme, and substrate mixture was added. After 10 minutes of room temperature pre-incubation, 10 μL of NADPH was added, and the plates were placed at 37° C. for 30 or 35 minutes depending on the enzyme being tested. After the incubation was complete, 20 μL of 0.1% Tris/ACN was added to the plate. The plate was read on a FluorStar fluorescence plate reader.
402
Total Activities
125
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Cytochrome P450 3A4 (CHEMBL340)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Inhibiting neurotransmitter reuptake
(2018)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 134 4.94 6.70
kon 134 - -
k_off 134 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL6010382 3 6.70
CHEMBL5982654 3 6.62
CHEMBL5943423 3 6.57
CHEMBL5939079 3 6.03
CHEMBL5866440 3 5.89
CHEMBL5870544 3 5.70
CHEMBL6055278 3 5.68
CHEMBL6015461 3 5.60
CHEMBL5875051 3 5.54
CHEMBL5769554 3 5.48
CHEMBL6004053 3 5.47
CHEMBL6011637 3 5.47
CHEMBL5865081 3 5.46
CHEMBL5904229 3 5.44
CHEMBL6048577 3 5.43
CHEMBL5994098 3 5.42
CHEMBL6006811 3 5.34
CHEMBL5955193 3 5.32
CHEMBL6055272 3 5.32
CHEMBL5777838 3 5.31
CHEMBL6063379 3 5.28
CHEMBL5739742 3 5.24
CHEMBL5905823 3 5.23
CHEMBL5769402 6 5.19
CHEMBL6032995 3 5.17
CHEMBL5831041 6 5.16
CHEMBL5920469 3 5.12
CHEMBL5800254 3 5.11
CHEMBL5748389 3 5.10
CHEMBL5773043 3 5.10

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL6010382 IC50 = 200.0 nM 6.70
CHEMBL5982654 IC50 = 240.0 nM 6.62
CHEMBL5943423 IC50 = 270.0 nM 6.57
CHEMBL5939079 IC50 = 940.0 nM 6.03
CHEMBL5866440 IC50 = 1300.0 nM 5.89
CHEMBL5870544 IC50 = 2000.0 nM 5.70
CHEMBL6055278 IC50 = 2100.0 nM 5.68
CHEMBL6015461 IC50 = 2500.0 nM 5.60
CHEMBL5875051 IC50 = 2900.0 nM 5.54
CHEMBL5769554 IC50 = 3300.0 nM 5.48
CHEMBL6004053 IC50 = 3400.0 nM 5.47
CHEMBL6011637 IC50 = 3400.0 nM 5.47
CHEMBL5865081 IC50 = 3500.0 nM 5.46
CHEMBL5904229 IC50 = 3600.0 nM 5.44
CHEMBL6048577 IC50 = 3700.0 nM 5.43
CHEMBL5994098 IC50 = 3800.0 nM 5.42
CHEMBL6006811 IC50 = 4600.0 nM 5.34
CHEMBL6055272 IC50 = 4800.0 nM 5.32
CHEMBL5955193 IC50 = 4800.0 nM 5.32
CHEMBL5777838 IC50 = 4900.0 nM 5.31
CHEMBL6063379 IC50 = 5300.0 nM 5.28
CHEMBL5739742 IC50 = 5700.0 nM 5.24
CHEMBL5905823 IC50 = 5900.0 nM 5.23
CHEMBL5769402 IC50 = 6400.0 nM 5.19
CHEMBL6032995 IC50 = 6700.0 nM 5.17
CHEMBL5831041 IC50 = 6900.0 nM 5.16
CHEMBL5920469 IC50 = 7600.0 nM 5.12
CHEMBL5800254 IC50 = 7800.0 nM 5.11
CHEMBL5748389 IC50 = 8000.0 nM 5.10
CHEMBL5773043 IC50 = 7900.0 nM 5.10
CHEMBL5882181 IC50 = 8100.0 nM 5.09
CHEMBL5812253 IC50 = 8200.0 nM 5.09
CHEMBL5798072 IC50 = 8100.0 nM 5.09
CHEMBL5852793 IC50 = 8300.0 nM 5.08
CHEMBL5893791 IC50 = 8400.0 nM 5.08
CHEMBL5929452 IC50 = 8600.0 nM 5.07
CHEMBL592374 AMITIFADINE IC50 = 8800.0 nM 5.06
CHEMBL5987298 IC50 = 8700.0 nM 5.06
CHEMBL5831041 IC50 = 9000.0 nM 5.05
CHEMBL5800266 IC50 = 9200.0 nM 5.04
CHEMBL5992805 IC50 = 9600.0 nM 5.02
CHEMBL6003284 IC50 = 9500.0 nM 5.02
CHEMBL5849289 IC50 = 9600.0 nM 5.02
CHEMBL5872179 IC50 = 10000.0 nM 5.00
CHEMBL5893791 IC50 = 9900.0 nM 5.00
CHEMBL5769402 IC50 = 11000.0 nM 4.96
CHEMBL5902505 IC50 = 11000.0 nM 4.96
CHEMBL5992805 IC50 = 11000.0 nM 4.96
CHEMBL5912067 IC50 = 11000.0 nM 4.96
CHEMBL5854009 IC50 = 13000.0 nM 4.89