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Assay Detail

CHEMBL3706280

Review assay metadata, readout intent, target linkage, and publication context from the same page.

Binding
Diluted TR-FRET Assay: TR-FRET AssayThe beta-secretase enzyme used in the TR-FRET is prepared as follows: The cDNA for the soluble part of the human beta-Secretase (AA 1-AA 460) was cloned using the ASP2-Fc10-1-IRES-GFP-neoK mammalian expression vector. The gene was fused to the Fc domain of IgG1 (affinity tag) and stably cloned into HEK 293 cells. Purified sBACE-Fc was stored in -80 C. in Tris buffer, pH 9.2 and had a purity of 40%.The enzyme (truncated form) was diluted to 6 ug/mL (stock 1.3 mg/mL) and the substrate (Europium)CEVNLDAEFK(Qsy7) to 200 nM (stock 120 uM) in reaction buffer (NaAcetate, chaps, triton x-100, EDTA pH4.5). The robotic systems Biomek FX and Velocity 11 were used for all liquid handling and the enzyme and substrate solutions were kept on ice until they were placed in the robotic system. Enzyme (9 ul) was added to the plate then 1 ul of compound in dimethylsulphoxide was added, mixed and pre-incubated for 10 minutes.
71
Total Activities
64
Compounds Tested
1
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Cell Type HEK293
Confidence 9 — Direct single protein target
Curated By Autocuration

Target

Beta-secretase 1 (CHEMBL4822)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Compounds and their use as BACE inhibitors
(2014)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 71 8.38 9.24

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3349234 1 9.24
CHEMBL3890867 1 9.20
CHEMBL3936953 2 9.15
CHEMBL3653702 1 9.15
CHEMBL3974225 1 9.14
CHEMBL3902302 1 9.08
CHEMBL2152917 1 9.05
CHEMBL3657429 1 9.00
CHEMBL3905878 1 8.96
CHEMBL3653723 2 8.92
CHEMBL3639562 1 8.89
CHEMBL3653726 1 8.89
CHEMBL2152919 1 8.85
CHEMBL2151058 1 8.85
CHEMBL3955716 1 8.82
CHEMBL3985518 1 8.82
CHEMBL2152915 1 8.80
CHEMBL2152918 1 8.80
CHEMBL3939410 1 8.68
CHEMBL3929159 1 8.66
CHEMBL2152914 1 8.66
CHEMBL2152916 2 8.66
CHEMBL3907727 1 8.66
CHEMBL3965844 1 8.64
CHEMBL3901045 1 8.64
CHEMBL2152920 1 8.64
CHEMBL3927408 1 8.60
CHEMBL3895159 1 8.59
CHEMBL3953357 1 8.55
CHEMBL3653722 2 8.54

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3349234 IC50 = 0.57 nM 9.24
CHEMBL3890867 IC50 = 0.63 nM 9.20
CHEMBL3936953 IC50 = 0.7 nM 9.15
CHEMBL3653702 IC50 = 0.7 nM 9.15
CHEMBL3974225 IC50 = 0.72 nM 9.14
CHEMBL3902302 IC50 = 0.84 nM 9.08
CHEMBL2152917 IC50 = 0.89 nM 9.05
CHEMBL3657429 IC50 = 1.0 nM 9.00
CHEMBL3905878 IC50 = 1.1 nM 8.96
CHEMBL3653723 IC50 = 1.2 nM 8.92
CHEMBL3653726 IC50 = 1.3 nM 8.89
CHEMBL3639562 IC50 = 1.3 nM 8.89
CHEMBL2151058 IC50 = 1.4 nM 8.85
CHEMBL2152919 IC50 = 1.4 nM 8.85
CHEMBL3955716 IC50 = 1.5 nM 8.82
CHEMBL3985518 IC50 = 1.5 nM 8.82
CHEMBL2152918 IC50 = 1.6 nM 8.80
CHEMBL2152915 IC50 = 1.6 nM 8.80
CHEMBL3936953 IC50 = 1.8 nM 8.74
CHEMBL3653723 IC50 = 2.0 nM 8.70
CHEMBL3939410 IC50 = 2.1 nM 8.68
CHEMBL2152916 IC50 = 2.2 nM 8.66
CHEMBL2152914 IC50 = 2.2 nM 8.66
CHEMBL3929159 IC50 = 2.2 nM 8.66
CHEMBL3907727 IC50 = 2.2 nM 8.66
CHEMBL3901045 IC50 = 2.3 nM 8.64
CHEMBL3965844 IC50 = 2.3 nM 8.64
CHEMBL2152920 IC50 = 2.3 nM 8.64
CHEMBL3927408 IC50 = 2.5 nM 8.60
CHEMBL3895159 IC50 = 2.6 nM 8.59
CHEMBL3953357 IC50 = 2.8 nM 8.55
CHEMBL3653722 IC50 = 2.9 nM 8.54
CHEMBL3657420 IC50 = 3.1 nM 8.51
CHEMBL3971150 IC50 = 3.1 nM 8.51
CHEMBL3894452 IC50 = 3.3 nM 8.48
CHEMBL3916726 IC50 = 3.3 nM 8.48
CHEMBL3923132 IC50 = 3.4 nM 8.47
CHEMBL3903281 IC50 = 3.9 nM 8.41
CHEMBL2152916 IC50 = 4.8 nM 8.32
CHEMBL3939422 IC50 = 4.9 nM 8.31
CHEMBL3653708 IC50 = 5.0 nM 8.30
CHEMBL3653697 IC50 = 5.3 nM 8.28
CHEMBL3972170 IC50 = 5.2 nM 8.28
CHEMBL3657419 IC50 = 5.3 nM 8.28
CHEMBL3653698 IC50 = 5.2 nM 8.28
CHEMBL3937468 IC50 = 5.5 nM 8.26
CHEMBL3653722 IC50 = 5.7 nM 8.24
CHEMBL3923132 IC50 = 5.7 nM 8.24
CHEMBL3902314 IC50 = 5.9 nM 8.23
CHEMBL3657420 IC50 = 6.1 nM 8.21