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Assay Detail

CHEMBL5737407

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Binding
Enzymatic Assay—1: The enzymatic assay couples DHODH activity with bleaching of the dye 2,6-dichlorophenolindophenol (DCIP) (Knecht and Loffler, 1998; Miller et al., 1968). The assay was conducted in buffer containing 50 mM Tris, 0.1% Triton X-100, 150 mM potassium chloride, 2 nM DHODH, 1 mM dihydroorotate, 0.1 mM decylubiquinone, 0.06 mM DCIP, and 2% DMSO at pH 8.0 at 32 degree celsius. The reaction was initiated by addition of substrates. Enzyme activity was monitored kinetically by the reduction in DCIP absorbance at 600 nm. Purified recombinant human DHODH enzyme was purchased from Novus (cat. no. NBP1-98916). Other chemicals were purchased from Sigma-Aldrich. Absorbance measurements were obtained using a BMG clarion star plate-reading spectrophotometer.
594
Total Activities
197
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

Dihydroorotate dehydrogenase (quinone), mitochondrial (CHEMBL1966)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

2,4,5-trisubstituted 1,2,4-triazolones useful as inhibitors of DHODH
(2021)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 198 7.17 9.00
kon 198 - -
k_off 198 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5821197 3 9.00
CHEMBL6028821 3 8.82
CHEMBL6038987 3 8.82
CHEMBL5791277 3 8.70
CHEMBL5899579 3 8.60
CHEMBL5936462 3 8.55
CHEMBL5871984 3 8.52
CHEMBL6040154 3 8.52
CHEMBL5880222 3 8.49
CHEMBL5945038 3 8.46
CHEMBL5961417 3 8.41
CHEMBL5077508 3 8.38
CHEMBL5855110 3 8.37
CHEMBL5843585 3 8.36
CHEMBL5818468 3 8.35
CHEMBL5881375 3 8.35
CHEMBL6002096 3 8.35
CHEMBL5894597 3 8.35
CHEMBL5943676 3 8.30
CHEMBL5926927 3 8.30
CHEMBL5837610 3 8.30
CHEMBL6011939 3 8.28
CHEMBL5793295 3 8.28
CHEMBL6049641 3 8.27
CHEMBL5979720 3 8.26
CHEMBL5837805 3 8.22
CHEMBL6034461 3 8.22
CHEMBL5765546 3 8.22
CHEMBL5852942 3 8.22
CHEMBL5834648 3 8.21

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5821197 IC50 = 1.0 nM 9.00
CHEMBL6038987 IC50 = 1.5 nM 8.82
CHEMBL6028821 IC50 = 1.5 nM 8.82
CHEMBL5791277 IC50 = 2.0 nM 8.70
CHEMBL5899579 IC50 = 2.5 nM 8.60
CHEMBL5936462 IC50 = 2.8 nM 8.55
CHEMBL6040154 IC50 = 3.0 nM 8.52
CHEMBL5871984 IC50 = 3.0 nM 8.52
CHEMBL5880222 IC50 = 3.2 nM 8.49
CHEMBL5945038 IC50 = 3.5 nM 8.46
CHEMBL5961417 IC50 = 3.9 nM 8.41
CHEMBL5077508 IC50 = 4.17 nM 8.38
CHEMBL5855110 IC50 = 4.3 nM 8.37
CHEMBL5843585 IC50 = 4.33 nM 8.36
CHEMBL5881375 IC50 = 4.5 nM 8.35
CHEMBL5894597 IC50 = 4.5 nM 8.35
CHEMBL5818468 IC50 = 4.5 nM 8.35
CHEMBL6002096 IC50 = 4.5 nM 8.35
CHEMBL5943676 IC50 = 5.0 nM 8.30
CHEMBL5926927 IC50 = 5.0 nM 8.30
CHEMBL5837610 IC50 = 5.0 nM 8.30
CHEMBL5793295 IC50 = 5.2 nM 8.28
CHEMBL6011939 IC50 = 5.2 nM 8.28
CHEMBL6049641 IC50 = 5.33 nM 8.27
CHEMBL5979720 IC50 = 5.5 nM 8.26
CHEMBL6034461 IC50 = 6.05 nM 8.22
CHEMBL5765546 IC50 = 6.0 nM 8.22
CHEMBL5837805 IC50 = 6.0 nM 8.22
CHEMBL5852942 IC50 = 6.0 nM 8.22
CHEMBL5834648 IC50 = 6.17 nM 8.21
CHEMBL6061058 IC50 = 6.5 nM 8.19
CHEMBL6054301 IC50 = 7.0 nM 8.15
CHEMBL5763638 IC50 = 7.0 nM 8.15
CHEMBL5900984 IC50 = 7.0 nM 8.15
CHEMBL5836077 IC50 = 7.5 nM 8.12
CHEMBL5984826 IC50 = 7.5 nM 8.12
CHEMBL5773494 IC50 = 7.5 nM 8.12
CHEMBL5939753 IC50 = 7.75 nM 8.11
CHEMBL5748647 IC50 = 8.5 nM 8.07
CHEMBL5910212 IC50 = 9.0 nM 8.05
CHEMBL5776168 IC50 = 9.75 nM 8.01
CHEMBL5842821 IC50 = 10.0 nM 8.00
CHEMBL5961667 IC50 = 11.0 nM 7.96
CHEMBL5918684 IC50 = 12.5 nM 7.90
CHEMBL5941952 IC50 = 13.0 nM 7.89
CHEMBL5973336 IC50 = 13.0 nM 7.89
CHEMBL5969863 IC50 = 13.0 nM 7.89
CHEMBL5817808 IC50 = 13.0 nM 7.89
CHEMBL5800827 IC50 = 13.1 nM 7.88
CHEMBL5771873 IC50 = 14.0 nM 7.85