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Assay Detail

CHEMBL5734901

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Binding
SMYD3 Biochemical Assay: The assays were all performed in a buffer consisting of 25 mM Tris-Cl pH 8.0, 1 mM TCEP, 0.005% BSG, and 0.005% Tween 20, prepared on the day of use. Compounds in 100% DMSO (1 ul) were spotted into a 384-well white opaque OptiPlate using a Bravo automated liquid handling platform outfitted with a 384-channel head (Agilent Technologies). DMSO (1 ul) was added to Columns 11, 12, 23, 24, rows A-H for the maximum signal control and 1 ul of SAH, a known product and inhibitor of SMYD3, was added to columns 11, 12, 23, 24, rows I-P for the minimum signal control. A cocktail (40 ul) containing the SMYD3 enzyme was added by Multidrop Combi (Thermo-Fisher). The compounds were allowed to incubate with SMYD3 for 30 min at room temperature, then a cocktail (10 ul) containing SAM and MEKK2 was added to initiate the reaction (final volume=51 ul). The final concentrations of the components were as follows: SMYD3 was 0.4 nM, 3H-SAM was 8 nM, MEKK2 was 12 nM, SAH in the minimum signal control wells was 1 mM, and the DMSO concentration was 2%. The assays were stopped by the addition of non-radiolabeled SAM (10 ul) to a final concentration of 100 uM, which dilutes the 3H-SAM to a level where its incorporation into MEKK2 is no longer detectable. Radiolabeled MEKK2 was detected using a scintillation proximity assay (SPA). 10 uL of a 10 mg/mL solution of SPA beads in 0.5 M citric acid was added and the plates centrifuged at 600 rpm for 1 min to precipitate the radiolabeled MEKK2 onto the SPA beads. The plates were then read in a PerkinElmer TopCount plate reader to measure the quantity of 3H-labeled MEKK2 as disintegrations per minute (dpm) or alternatively, referred to as counts per minute (cpm).% Inhibition Calculation% ⁢ ⁢ inh = 100 - ( dpm cmpd - dpm min dpm max - dpm min ) × 100Where dpm=disintegrations per minute, cmpd=signal in assay well, and min and max are the respective minimum and maximum signal controls.Four-Parameter IC50 FitY = Bottom + ( Top - Bottom ) ( 1 + ( X IC 50 ) Hill ⁢ ⁢ CoefficientWhere top and bottom are the normally allowed to float, but may be fixed at 100 or 0 respectively in a 3-parameter fit. The Hill Coefficient normally allowed to float but may also be fixed at 1 in a 3-parameter fit. Y is the % inhibition and X is the compound concentration.
ChEMBL ↗
2955
Total Activities
978
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Confidence 8 — Homologous single protein target
Curated By Autocuration

Target

Histone-lysine N-methyltransferase SMYD3 (CHEMBL2321643) ↗
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Substituted piperidine compounds
(2020)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 985 6.82 9.40
kon 985 - -
k_off 985 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5930708 — — 3 9.40
CHEMBL6062390 — — 3 9.22
CHEMBL5915469 — — 3 9.15
CHEMBL5817493 — — 3 9.12
CHEMBL5983928 — — 3 9.10
CHEMBL5749196 — — 3 9.10
CHEMBL5976516 — — 3 9.06
CHEMBL5767419 — — 3 9.05
CHEMBL5964919 — — 3 9.05
CHEMBL5956627 — — 3 9.05
CHEMBL5777063 — — 3 9.03
CHEMBL5759159 — — 3 9.02
CHEMBL5880073 — — 6 9.00
CHEMBL5976948 — — 3 9.00
CHEMBL5989847 — — 3 8.99
CHEMBL6054359 — — 3 8.98
CHEMBL5910850 — — 3 8.97
CHEMBL6006815 — — 3 8.96
CHEMBL5874113 — — 3 8.95
CHEMBL6040217 — — 3 8.94
CHEMBL5874030 — — 3 8.93
CHEMBL5920563 — — 3 8.92
CHEMBL5847089 — — 3 8.92
CHEMBL5984539 — — 3 8.92
CHEMBL5843540 — — 3 8.92
CHEMBL6020373 — — 3 8.91
CHEMBL5777827 — — 3 8.91
CHEMBL5845495 — — 3 8.91
CHEMBL5887443 — — 3 8.89
CHEMBL5894057 — — 3 8.89

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5930708 — IC50 = 0.4 nM 9.40
CHEMBL6062390 — IC50 = 0.6 nM 9.22
CHEMBL5915469 — IC50 = 0.71 nM 9.15
CHEMBL5817493 — IC50 = 0.76 nM 9.12
CHEMBL5983928 — IC50 = 0.8 nM 9.10
CHEMBL5749196 — IC50 = 0.8 nM 9.10
CHEMBL5976516 — IC50 = 0.88 nM 9.06
CHEMBL5767419 — IC50 = 0.9 nM 9.05
CHEMBL5964919 — IC50 = 0.9 nM 9.05
CHEMBL5956627 — IC50 = 0.9 nM 9.05
CHEMBL5777063 — IC50 = 0.94 nM 9.03
CHEMBL5759159 — IC50 = 0.95 nM 9.02
CHEMBL5976948 — IC50 = 1.0 nM 9.00
CHEMBL5880073 — IC50 = 1.0 nM 9.00
CHEMBL5989847 — IC50 = 1.03 nM 8.99
CHEMBL6054359 — IC50 = 1.04 nM 8.98
CHEMBL5910850 — IC50 = 1.07 nM 8.97
CHEMBL6006815 — IC50 = 1.1 nM 8.96
CHEMBL5874113 — IC50 = 1.12 nM 8.95
CHEMBL6040217 — IC50 = 1.14 nM 8.94
CHEMBL5874030 — IC50 = 1.18 nM 8.93
CHEMBL5843540 — IC50 = 1.21 nM 8.92
CHEMBL5984539 — IC50 = 1.2 nM 8.92
CHEMBL5920563 — IC50 = 1.2 nM 8.92
CHEMBL5847089 — IC50 = 1.2 nM 8.92
CHEMBL5845495 — IC50 = 1.24 nM 8.91
CHEMBL6020373 — IC50 = 1.22 nM 8.91
CHEMBL5777827 — IC50 = 1.23 nM 8.91
CHEMBL5966075 — IC50 = 1.29 nM 8.89
CHEMBL5887443 — IC50 = 1.3 nM 8.89
CHEMBL5894057 — IC50 = 1.3 nM 8.89
CHEMBL5678791 — IC50 = 1.3 nM 8.89
CHEMBL5917319 — IC50 = 1.3 nM 8.89
CHEMBL5930979 — IC50 = 1.36 nM 8.87
CHEMBL5818034 — IC50 = 1.37 nM 8.86
CHEMBL5753505 — IC50 = 1.38 nM 8.86
CHEMBL5918993 — IC50 = 1.4 nM 8.85
CHEMBL5792955 — IC50 = 1.42 nM 8.85
CHEMBL5961592 — IC50 = 1.59 nM 8.80
CHEMBL5795246 — IC50 = 1.6 nM 8.80
CHEMBL5880073 — IC50 = 1.8 nM 8.74
CHEMBL5962050 — IC50 = 1.81 nM 8.74
CHEMBL5951905 — IC50 = 1.82 nM 8.74
CHEMBL5941012 — IC50 = 1.85 nM 8.73
CHEMBL5836468 — IC50 = 1.87 nM 8.73
CHEMBL5975793 — IC50 = 1.87 nM 8.73
CHEMBL5998284 — IC50 = 1.85 nM 8.73
CHEMBL5768086 — IC50 = 1.9 nM 8.72
CHEMBL5871188 — IC50 = 1.9 nM 8.72
CHEMBL5744078 — IC50 = 1.9 nM 8.72