## Endpoint to Predict
- Endpoint scope: assay
- Measurement type: IC50
- Label type: continuous
- Binary semantics: pchembl
- Assay description: Assay on In Vitro Enzyme-Inhibiting Activity: Test compounds: a part of compounds of the invention, the chemical names and preparation methods of which can be found in their preparation examples.Control agent: CO-1686, the structure of which can be found in the Background Art, prepared by the inventors (please refer to Patent WO2012061299A1 for the preparation methods).The meanings represented by the abbreviations in the experiments are described as follows.EDTA: eathylene diamine tetraacetic acidDMSO: dimethyl sulfoxideSD: standard deviationFAM: carboxyfluoresceinBrij-35: polyethylene glycol dodecyl etherHEPES: N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acidDTT: dithiothreitolExperimental method: the compounds were screened in the presence of Km ATP by Mobility Shift Assay using the kinases EGFR and EGFR_T790M.1. Preparation of Reagents(1) 1-fold kinase buffer for detecting the kinases wild-type EGFR (WT EGFR or WT for short), EGFR (d746-750) (d746-750 for short), EGFR (d746-750)-T790M ((d746-750)-T790M for short): 50 mM HEPES (pH 7.5), 0% Brij-35, 10 mM MgCl2, 10 mM MnCl2, and 2 mM DTT;(2) 1-fold kinase buffer for detecting the kinases EGFR T790M (T790M for short), EGFR L858R (L858R for short), EGFR T790M-L858R (T790M-L858R for short): 50 mM HEPES (pH 7.5), 0% Brij-35, 5 mM MgCl2, and 2 mM DTT.(3) Stop Solution100 mM HEPES (pH 7.5), 0.01% Brij-35, 0.2% Coating Reagent #3, and 50 mM EDTA2. Preparation of a Compound Solution(1) To the second well of a 96-well plate, 5 μL 10 mM compound (dissolved in DMSO) was added, and 95 μL 100% DMSO was added to prepare 100 μL 0.5 mM compound.(2) To other wells, 60 μL 100% DMSO was added. 20 μL compound from the second well was added to the third well, and 4-fold dilution was further performed to get 10 diluted concentrations.(3) To the first well and the twelfth well of the 96-well plate, 100 μL 100% DMSO was separately added, and the two wells were used as control wells.(4) 10 μL from each well of the 96-well plate was added to another 96-well plate, and 90 μL 1-fold kinase buffer was added.(5) 5 μL from the 96-well plate was added to another 384-well plate, for example, transferred from A1 well of the 96-well plate to A1 and A2 wells of the 384-well plate, and transferred from A2 well of the 96-well plate to A3 and A4 wells of the 384-well plate, and so on.3. Kinase reaction(1) Preparation of 2.5-fold enzyme solutionKinase was added to 1-fold kinase buffer to form 2.5-fold enzyme solution.(2) Preparation of 2.5-fold substrate solutionFAM-tagged polypeptide and ATP were added to 1-fold kinase buffer to form 2.5-fold substrate solution.(3) Addition of 2.5-fold enzyme solution to 384-well plateTo 384-well plate, 10 μL 2.5-fold enzyme solution was added, and incubated at room temperature for 10 min.(4) Addition of 2.5-fold substrate solution to 384-well plateTo 384-well plate, 10 μL 2.5-fold substrate solution was added.(5) Kinase reaction and stopAfter incubation at 28° C. for a period of time (depending on kinase), 25 μL stop solution was added.4. Data on percent conversion was read by Caliper.5. Data analysis(1) Data on enzyme activity was obtained by Caliper program;(2) Inhibition rate of enzyme activity was calculated from the data on enzyme activity by the following formula: Inhibition rate(%)=(maximal value−the measured value of a test compound)/(maximal value-minimal value)×100,wherein, the maximal value represents the measured value of DMSO control; and the minimal value represents the measured value of blank control.(3) Calculation of IC50 value by XLFit excel.
- Target name: Epidermal growth factor receptor
- Target type: SINGLE PROTEIN
- Target organism: Homo sapiens

## Reference Molecule A
- Canonical SMILES: C=CC(=O)Nc1cccc(Nc2nc(Nc3ccc(N4CCC(N5CCOCC5)CC4)cc3OC)ncc2C(F)(F)F)c1
- Properties:
- ChEMBL ID: CHEMBL5767529
- Molecular weight: 597.64
- Heavy atoms: 43
- Hydrogen bond donors: 3
- Hydrogen bond acceptors: 9
- TPSA: 103.88
- LogP: 5.42
- Rotatable bonds: 9
- Ring count: 5
- Aromatic rings: 3
- Formal charge: 0
- Fraction Csp3: 0.37
- QED: 0.28

## Known Reference Result on This Endpoint
- Molecule A clean result: median pChEMBL = 9.1
- This result is provided as context for predicting Molecule B on the same endpoint.

## Reference Molecule A Other ChEMBL Evidence
- type IC50; relation =; value 0.8; units nM; pChEMBL 9.1; comment 782784

## Query Molecule B
- Canonical SMILES: C=CC(=O)Nc1cccc(Nc2nc(Nc3ccc(C4CCN(S(C)(=O)=O)CC4)cc3OC)ncc2C(F)(F)F)c1
- Properties:
- ChEMBL ID: CHEMBL6049383
- Molecular weight: 590.63
- Heavy atoms: 41
- Hydrogen bond donors: 3
- Hydrogen bond acceptors: 8
- TPSA: 125.55
- LogP: 5.25
- Rotatable bonds: 9
- Ring count: 4
- Aromatic rings: 3
- Formal charge: 0
- Fraction Csp3: 0.3
- QED: 0.29

## Molecule Similarity
- Weighted Tanimoto: 0.64

Answer: 8.44