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Assay Detail

CHEMBL3885776

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

Binding
Biochemical assay using fluorogenic peptide substrate.Step 1: Compound transfer to plate (50 nl).Step 2: Dilute FXa in Assay Buffer.Step 3: Dilute substrate in Assay Buffer.Step 4: Add equal volumes to Assay plate (total assay volume 10 uL).Step 5: Centrifuge plate, 1 min, 1000 rpm.Step 6: Detect fluorescence signal: ex: 320 nm; Em. 405 nm (EnVision), every 10 min. 0 min pre-incubation of enzyme with compound and 30 min reaction time after substrate addition.
ChEMBL ↗
54
Total Activities
54
Compounds Tested
1
Activity Types
20
Assay Parameters

Assay Information

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

Target

Coagulation factor X (CHEMBL244) ↗
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

K4DD drug target binding kinetics data
Doris A.Schuetz, Wilhelmus Egbertus Arnout de Witte, Yin Cheong Wong, Bernhard Knasmueller, Lars Richter, Daria B. Kokh, S. Kashif Sadiq, Reggie Bosma, Indira Nederpelt, Laura H. Heitman, Elena Segala, Marta Amaral, Dong Guo, Dorothee Andres, Victoria Georgi, Leigh A. Stoddart, Steve Hill, Robert M. Cooke, Chris De Graaf, Rob Leurs, Matthias Frech, Rebecca C. Wade, Elizabeth Cunera Mariade Lange, Adriaan P. IJzerman, Anke Müller-Fahrnow, Gerhard F. Ecker

· DOI

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 54 8.13 9.69

Assay Parameters

Parameter Type Value Units Text
assay_comment assay_comment - — Assay Buffer: 50 mM Tris, 5 mM CaCl2, 150 mM NaCl, 0.05 % Brij-35, pH 7.5. Assay plate: Black, 384 well plate, small volume, non-biniding, Greiner Bio-One (784900). For full SOP, see SOP-IME-SP-023 Biochemical FI Assay for FXa_v01.pdf.
assay_method assay_method - — Biochemical
biochemical_mode biochemical_mode - — direct binding
expression_host expression_host - — Insect cells
fitting_model fitting_model - — 4-parameter logistic fit (for IC50 determinations)
incubation_time incubation_time 30.0 min —
internal_assay_id internal_assay_id - — SOP-IME-SP-023
Km Km 56700.0 nM —
number_control_wells number_control_wells - — 16 negative controls and 16 positive controls (50µM PPACK)
plate_layout plate_layout - — 16x24
pre_incubation_time pre_incubation_time - min —
protein_concentration protein_concentration 0.0025 uM —
protein_molecular_weight protein_molecular_weight 46.6 kDa —
protein_mutation_type protein_mutation_type - — native protein
reference_substance reference_substance - — D-Phe-Pro-Arg-CMK (PPACK; N-1065 from BACHEM)
sample_buffer_composition sample_buffer_composition - — 50 mM Tris, 5 mM CaCl2, 150 mM NaCl, 0.05 % Brij-35
sample_ph sample_ph 7.5 — —
sample_temperature sample_temperature 25.0 degree C —
signal_stability signal_stability 1.0 — —
s_b_ratio s_b_ratio 2.0 — —

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL364284 — — 1 9.69
CHEMBL197377 — — 1 9.27
CHEMBL3960154 — — 1 9.22
CHEMBL46618 OTAMIXABAN 3.0 1 9.15
CHEMBL370614 — — 1 9.15
CHEMBL3961484 — — 1 9.12
CHEMBL3910588 — — 1 9.09
CHEMBL3972799 — — 1 9.09
CHEMBL194186 — — 1 9.08
CHEMBL196539 — — 1 9.08
CHEMBL383361 — — 1 9.07
CHEMBL3986101 — — 1 8.94
CHEMBL3979386 — — 1 8.94
CHEMBL564248 — — 1 8.90
CHEMBL48813 — — 1 8.81
CHEMBL3912108 — — 1 8.80
CHEMBL190142 — — 1 8.77
CHEMBL557915 — — 1 8.76
CHEMBL3904655 — — 1 8.74
CHEMBL3907419 — — 1 8.72
CHEMBL3955803 — — 1 8.71
CHEMBL372956 — — 1 8.69
CHEMBL3968842 — — 1 8.66
CHEMBL3958789 — — 1 8.57
CHEMBL381866 — — 1 8.44
CHEMBL196162 — — 1 8.41
CHEMBL198654 — — 1 8.36
CHEMBL196492 — — 1 8.32
CHEMBL3304291 — — 1 8.28
CHEMBL550453 — — 1 8.23

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL364284 — IC50 = 0.203 nM 9.69
CHEMBL197377 — IC50 = 0.542 nM 9.27
CHEMBL3960154 — IC50 = 0.599 nM 9.22
CHEMBL46618 OTAMIXABAN IC50 = 0.702 nM 9.15
CHEMBL370614 — IC50 = 0.707 nM 9.15
CHEMBL3961484 — IC50 = 0.767 nM 9.12
CHEMBL3910588 — IC50 = 0.816 nM 9.09
CHEMBL3972799 — IC50 = 0.813 nM 9.09
CHEMBL194186 — IC50 = 0.835 nM 9.08
CHEMBL196539 — IC50 = 0.824 nM 9.08
CHEMBL383361 — IC50 = 0.853 nM 9.07
CHEMBL3986101 — IC50 = 1.16 nM 8.94
CHEMBL3979386 — IC50 = 1.16 nM 8.94
CHEMBL564248 — IC50 = 1.26 nM 8.90
CHEMBL48813 — IC50 = 1.54 nM 8.81
CHEMBL3912108 — IC50 = 1.59 nM 8.80
CHEMBL190142 — IC50 = 1.7 nM 8.77
CHEMBL557915 — IC50 = 1.73 nM 8.76
CHEMBL3904655 — IC50 = 1.8 nM 8.74
CHEMBL3907419 — IC50 = 1.91 nM 8.72
CHEMBL3955803 — IC50 = 1.94 nM 8.71
CHEMBL372956 — IC50 = 2.03 nM 8.69
CHEMBL3968842 — IC50 = 2.17 nM 8.66
CHEMBL3958789 — IC50 = 2.68 nM 8.57
CHEMBL381866 — IC50 = 3.67 nM 8.44
CHEMBL196162 — IC50 = 3.87 nM 8.41
CHEMBL198654 — IC50 = 4.39 nM 8.36
CHEMBL196492 — IC50 = 4.77 nM 8.32
CHEMBL3304291 — IC50 = 5.23 nM 8.28
CHEMBL550453 — IC50 = 5.82 nM 8.23
CHEMBL3972969 — IC50 = 6.15 nM 8.21
CHEMBL3956658 — IC50 = 6.55 nM 8.18
CHEMBL196551 — IC50 = 7.33 nM 8.13
CHEMBL3957468 — IC50 = 12.7 nM 7.90
CHEMBL194810 — IC50 = 13.7 nM 7.86
CHEMBL3974641 — IC50 = 18.0 nM 7.75
CHEMBL3305961 — IC50 = 18.1 nM 7.74
CHEMBL1235423 — IC50 = 21.8 nM 7.66
CHEMBL371106 — IC50 = 31.9 nM 7.50
CHEMBL316053 — IC50 = 32.4 nM 7.49
CHEMBL197547 — IC50 = 36.3 nM 7.44
CHEMBL197624 — IC50 = 38.1 nM 7.42
CHEMBL3959350 — IC50 = 38.5 nM 7.42
CHEMBL3928201 — IC50 = 38.6 nM 7.41
CHEMBL93087 — IC50 = 57.0 nM 7.24
CHEMBL120413 — IC50 = 58.5 nM 7.23
CHEMBL436293 — IC50 = 87.0 nM 7.06
CHEMBL196589 — IC50 = 124.0 nM 6.91
CHEMBL370176 — IC50 = 152.0 nM 6.82
CHEMBL198421 — IC50 = 250.0 nM 6.60