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Assay Detail

CHEMBL5736349

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Binding
Fluorescence Polarization (FP) Competitive Binding Assay: A fluorescence polarization (FP) competitive binding assay was used to determine the binding affinities of representative menin inhibitors. A FAM labeled fluorescent probe was designed and synthesized based on a MLL1 peptide (FAM-MM2). Equilibrium dissociation constant (Kd) value of FAM-MM2 to menin protein was determined from protein saturation experiments by monitoring the total fluorescence polarization of mixtures composed with the fluorescent probe at a fixed concentration and the protein with increasing concentrations up to full saturation. Serial dilutions of the protein were mixed with FAM-MM2 to a final volume of 200 μl in the assay buffer (PBS with 0.02% Bovine γ-Globulin and 4% DMSO. 0.01% Triton X-100 was added right before assays). Final FAM-MM2 concentration was 2 nM. Plates were incubated at room temperature for 30 minutes with gentle shaking to assure equilibrium. FP values in millipolarization units (mP) were measured using the Infinite M-1000 plate reader (Tecan U.S., Research Triangle Park, N.C.) in Microfluor 1 96-well, black, v-bottom plates (Thermo Scientific, Waltham, Mash.) at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. Kd value of FAM-MM2, which was calculated by fitting the sigmoidal dose-dependent FP increases as a function of protein concentrations using Graphpad Prism 6.0 software (Graphpad Software, San Diego, Calif.), is determined as 1.4 nM.
1158
Total Activities
378
Compounds Tested
3
Activity Types
0
Assay Parameters

Assay Information

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

Target

Menin (CHEMBL1615381)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Piperidines as menin inhibitors
(2021)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 386 6.53 8.52
kon 386 - -
k_off 386 - -

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL5816906 3 8.52
CHEMBL5974610 3 8.52
CHEMBL5905647 3 8.52
CHEMBL5931080 3 8.52
CHEMBL5964939 3 8.52
CHEMBL6047118 3 8.52
CHEMBL5798441 3 8.40
CHEMBL5829566 3 8.40
CHEMBL5938942 3 8.40
CHEMBL5975630 3 8.40
CHEMBL5822575 3 8.40
CHEMBL5985246 3 8.40
CHEMBL5743156 3 8.40
CHEMBL6020537 3 8.30
CHEMBL5948342 3 8.30
CHEMBL5960556 3 8.30
CHEMBL6013737 6 8.30
CHEMBL4585198 3 8.22
CHEMBL5790028 3 8.22
CHEMBL5783513 3 8.22
CHEMBL5767175 3 8.22
CHEMBL6034357 3 8.22
CHEMBL5893927 3 8.22
CHEMBL4551865 3 8.15
CHEMBL4592636 3 8.15
CHEMBL5843849 3 8.15
CHEMBL5769793 3 8.15
CHEMBL5777937 3 8.15
CHEMBL5859938 3 8.15
CHEMBL5783978 3 8.15

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL5905647 IC50 = 3.0 nM 8.52
CHEMBL5931080 IC50 = 3.0 nM 8.52
CHEMBL5816906 IC50 = 3.0 nM 8.52
CHEMBL5974610 IC50 = 3.0 nM 8.52
CHEMBL5964939 IC50 = 3.0 nM 8.52
CHEMBL6047118 IC50 = 3.0 nM 8.52
CHEMBL5743156 IC50 = 4.0 nM 8.40
CHEMBL5798441 IC50 = 4.0 nM 8.40
CHEMBL5938942 IC50 = 4.0 nM 8.40
CHEMBL5822575 IC50 = 4.0 nM 8.40
CHEMBL5985246 IC50 = 4.0 nM 8.40
CHEMBL5829566 IC50 = 4.0 nM 8.40
CHEMBL5975630 IC50 = 4.0 nM 8.40
CHEMBL6013737 IC50 = 5.0 nM 8.30
CHEMBL5948342 IC50 = 5.0 nM 8.30
CHEMBL5960556 IC50 = 5.0 nM 8.30
CHEMBL6020537 IC50 = 5.0 nM 8.30
CHEMBL5790028 IC50 = 6.0 nM 8.22
CHEMBL4585198 IC50 = 6.0 nM 8.22
CHEMBL6034357 IC50 = 6.0 nM 8.22
CHEMBL5783513 IC50 = 6.0 nM 8.22
CHEMBL5767175 IC50 = 6.0 nM 8.22
CHEMBL5893927 IC50 = 6.0 nM 8.22
CHEMBL4551865 IC50 = 7.0 nM 8.15
CHEMBL5859938 IC50 = 7.0 nM 8.15
CHEMBL5777937 IC50 = 7.0 nM 8.15
CHEMBL5769793 IC50 = 7.0 nM 8.15
CHEMBL5935610 IC50 = 7.0 nM 8.15
CHEMBL5783978 IC50 = 7.0 nM 8.15
CHEMBL4592636 IC50 = 7.0 nM 8.15
CHEMBL5899857 IC50 = 7.0 nM 8.15
CHEMBL5843849 IC50 = 7.0 nM 8.15
CHEMBL5936667 IC50 = 8.0 nM 8.10
CHEMBL5962223 IC50 = 8.0 nM 8.10
CHEMBL5788808 IC50 = 8.0 nM 8.10
CHEMBL5893826 IC50 = 8.0 nM 8.10
CHEMBL6029944 IC50 = 8.0 nM 8.10
CHEMBL5769865 IC50 = 8.0 nM 8.10
CHEMBL5836021 IC50 = 8.0 nM 8.10
CHEMBL5820786 IC50 = 8.0 nM 8.10
CHEMBL5848603 IC50 = 8.0 nM 8.10
CHEMBL4449199 IC50 = 8.0 nM 8.10
CHEMBL5835328 IC50 = 9.0 nM 8.05
CHEMBL5794080 IC50 = 9.0 nM 8.05
CHEMBL4522767 IC50 = 9.0 nM 8.05
CHEMBL5865073 IC50 = 9.0 nM 8.05
CHEMBL5768069 IC50 = 9.0 nM 8.05
CHEMBL5765474 IC50 = 9.0 nM 8.05
CHEMBL5970007 IC50 = 9.0 nM 8.05
CHEMBL5740449 IC50 = 9.0 nM 8.05