External ID: CHEMBL1266185

Protocol: N/A

Comment: Journal: Nat Chem Biol
Year: 2007
Volume: 3
Issue: 5
First Page: 268
Last Page: 273
DOI: 10.1038/nchembio873

External ID: SNCA-p-activity-luciferase

Protocol: PROTOCOL TABLE
SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE; DESCRIPTION

1; Cells; 4 uL; Dispense 1500 HEK-293-SNCA-luc cells/well into Greiner 1536-well white / solid bottom tissue culture treated plate. The plate was covered with metal lids with gas-exchange holes.
2; Incubate; 24 hours; Incubate at 37C, 5% CO2, 95% RH.
3; Compounds; 23 nL; Compounds and controls were transferred via a Kalypsys Pin Tool (Wako USA) equipped with a 1536-slotted pin array. The plate was covered with metal lids with gas-exchange holes.
4; Incubate; 24 hours; Incubate at 37C, 5% CO2, 95% RH.
5; Dispense; 1 uL; Dispense Gly-Phe-7-amino-4-trifluoromethylcoumarin (GF-AFC, prepared at 125 uM in PBS) was added. The plate was covered with metal lids with gas-exchange holes.
6; Incubate; 30 min; Incubate at 37C, 5% CO2.
7; Detector; Fluorescence; Measure fluorescence with ViewLux microplate reader (PerkinElmer) equipped with 405/10 excitation and 540/25 emission filters.
8; Dispense; 3 uL; Dispense ONE-Glo (PerkinElmer) lucifase detection reagent was added to each well. Plates were covered with metal lids with gas-exchange holes.
9; Incubate; 15 min; Incubate at room temperature.
10; Detector; Luminescence; Measure luminescence with ViewLux microplate reader (PerkinElmer) equipped with clear filters.

NOTES (numbers refer to sequence above)
1; HEK-293-SNCA-luc were cultured and suspended in phenol-red free DMEM (4.5 g/L glucose, 25 mM HEPES, cat #21063 (Thermo)).
3; Compounds were added to the assay plate in an 11-point intra plate dose response, 1:3 titration in DMSO with a final concentration range of xxx - yyy uM. Vehicle-only plates, with DMSO being pin-transferred to every well, were inserted at the beginning of screening runs to confirm expected assay performance. Activity was normalized to wells containing medium only (-100% activity, full inhibition) and SNCA-luc cells treated with DMSO vehicle control (0% activity), contained on the same plate as test samples.
10; Signals were analyzed, and dose-response curves were fit using the Hill equation. Compounds in curve classes -1.1, -1.2, -2.1, -2.2 in the SNCA-luc assay were considered active. Compounds were eliminated from further consideration if also active (curve class -1.1, -1.2, -1.3, -1.4, -2.1, -2.2, -2.3, -2.4) in the GF-AFC cytotoxicity assay.

Comment: Disclaimer:
Although all reasonable efforts have been made to ensure the accuracy and reliability of the data, caution should be exercised when interpreting the results as artifacts are possible from nonspecific effects such as assay signal interference. The curve fitting and activity calls presented here are based on the NCATS analysis methods.

Compound Ranking:
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, with a ratio activity curve class of 4, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. For a ratio activity curve class = -1.1, score = 80+abs((log_ac50+4.5)*inf_activity/20). For ratio.curve_class == -1.2 && abs(ratio.max_response) > 6*10, score = 60+abs((log_ac50+4.5)*inf_activity/20). For ratio.curve_class == -2.1 || ( ratio.curve_class==-2.2 && abs(ratio.max_response) > 6*10), score = 40+abs((log_ac50+4.5)*inf_activity/20). For ratio.curve_class == -1.2 || ratio.curve_class == -2.2, score = 20+abs((log_ac50+4.5)*inf_activity/20). Inconclusive compounds, with a donor curve class other than 4, have PUBCHEM_ACTIVITY_SCORE of 10. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39.

External ID: CHEMBL1220301

Protocol: N/A

Comment: Journal: Nat. Chem. Biol.
Year: 2008
Volume: 4
Issue: 3
First Page: 197
Last Page: 199
DOI: 10.1038/nchembio.65

Target ChEMBL ID: CHEMBL3308902
ChEMBL Target Name: Eukaryotic peptide chain release factor GTP-binding subunit
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: CHEMBL4709071

Protocol: N/A

Comment: Compounds with activity <= 10uM or explicitly reported as active by ChEMBL are flagged as active in this PubChem assay presentation.

Journal: Eur J Med Chem
Year: 2020
Volume: 204
First Page: 112505
Last Page: 112505
DOI: 10.1016/j.ejmech.2020.112505

Target ChEMBL ID: CHEMBL5026
ChEMBL Target Name: Serine/threonine-protein kinase EEF2K
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: TargetID_659_CEMA

Protocol: Black, standard capacity streptavidin-coated 384-well plates (Pierce 15407) were first washed with 50 L of phosphate buffer (100 mM, pH 7.0; PB7) three times using a Biotek 405 ELX plate washer. Subsequently, 5 L of biotinylated pre-miRNA substrate (500 nM final) was dispensed into the plate using a Multidrop Combi Reagent Dispenser (Thermo Scientific). Plates were then centrifuged for 1 min at 1,000 RPM (223 g), sealed with plate tape, and incubated overnight at 4 C. The following morning, plates were washed three times with 50 L of PB7, followed by the addition of 5 L of Dicer digest buffer (20 mM Tris, 12 mM NaCl, 2.5 mM MgCl2, 1 mM fresh DTT, and 4.5% DMSO) and centrifugation. Compounds (50 nL of 5 mM DMSO stock, 25 M final) were then added into the sample wells using a Sciclone (Caliper) liquid handler with V&P pintool; the same volume of DMSO was added to the control wells. The plates were incubated at 25 C for 15 min before addition of 5 L of digest buffer containing 217 g/nL Dicer (108 g/mL Dicer, 5% glycerol and 0.01% Triton X-100 final, excess with respect to pre-miRNA). For the positive control wells, digest buffer without Dicer was added. The plates were centrifuged again and resealed before being placed in a 37 C incubator for 5 h. After Dicer cleavage, plates were washed three times with 50 L of PB7. mTet-HRP in PB7 (10 L, 750 nM final) was then dispensed into each well. The plates were subsequently centrifuged, sealed, and incubated at 25 C for 2 h. Plates were then washed three times with 50 L of wash buffer (2 mM imidazole, 260 mM NaCl, 0.5 mM EDTA, 0.1% Tween-20, pH 7.0), followed by washing three additional times with 50 L of PB7. Finally, SuperSignal West Pico (25 L; Pierce) was added, the plates were incubated at 25 C for 5 min, and chemiluminescence signal was detected using a PHERAstar plate reader using LUM plus module (BMG Labtech).

Comment: The activity outcome is based on a Z-score (number of standard deviations from the negative control mean) of 3 or higher on at least 50% times that the sample was screened.

For instance, if the sample was screened in n=4 runs, it would be considered active only if it had a Z-score of 3 or above in at least 2 runs.

External ID: CHEMBL1931128

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2012
Volume: 47
First Page: 214
Last Page: 220
DOI: 10.1016/j.ejmech.2011.10.047

Target ChEMBL ID: CHEMBL2996
ChEMBL Target Name: Protein kinase C delta
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: CHEMBL1931129

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2012
Volume: 47
First Page: 214
Last Page: 220
DOI: 10.1016/j.ejmech.2011.10.047

Target ChEMBL ID: CHEMBL6052
ChEMBL Target Name: Transforming protein RhoA
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: CHEMBL2429086

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429085

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429084

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429083

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429090

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429089

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429088

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429087

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: HERG01

Protocol: NCGC Assay Protocol Summary:

HERG assay (FluxORTM thallium flux assay) was initially developed by Invitrogen/Molecular Probes, and then miniaturized into 1536-well plate in a homogeneous format by NCGC. This assay measures the activity of potassium channel using thallium dye (FluxOR) flux as surrogate measurement for potassium into the cells with a FDSS-7000 kinetic plate reader (Hamamatsu Corp., Hamamatsu City, Japan). The hERG ion channel is transduced into mammalian cells (U2OS) using a baculovirus (Bacmam) construct harboring the hERG K+ ion channel. So far we screened LOPAC1280 library (Sigma), the NTP collection of 1408 compounds, and the NCGC Pharmaceutical Collection (NPC), in which many well defined HERG blockers are present. The rank order potencies of many of these compounds are similar to that of other HERG assays (membrane potential, Rb+ flux, patch clamp, etc). This quick and homogeneous assay is also found to be sensitive, specific, and robust.

Using the FluxORTM thallium flux assay, the activity of potassium channel using thallium dye (FluxOR) flux as surrogate measurement for potassium into the cells was measured in the U2OS cell line transduced with hERG K+ ion channel using a baculovirus (Bacmam) using Opti-MEM medium (Invitrogen) containing 2% fetal calf serum (FCS, HyCone) following loading buffer addition, compound treatment for around 10 minutes and finally adding stimulation buffer. The assay was performed in black clear Kalypsys 1536-well plates. In the screen, Astemizole was used as positive controls. Library compounds were measured for their ability to cause hERG channel blockage in the cell line, as reflected by a decrease in fluorescence intensity, in a concentration-dependent manner. Data were normalized to the controls for basal activity (DMSO only) and 100% inhibition (5uM Astemizole). AC50 values were determined from concentration-response data modeled with the standard Hill equation.

qHTS protocol for hERG-U2OS cellular assay

[Step] [Parameter] [Value] [Description]

1. Day 1: Replace medium in 70-80% confluent T225 flask with 2.5 mL of hERG-BacMam virus plus 12.5 mL of phosphate buffered saline (PBS) (corresponding roughly to a multiplicity of infection ratio of 100 virus particles/cell)
2. Incubation: 4 hrs @ room Temperature in Dark
3. Reagent; Remove virus; wash once with 25 ml DPBS
4. Reagent; 35 ml culture medium
5. Incubation; 37oC overnight
6. Day2: Reagent; 3 uL; 2000 U2OS cells/well
7. Time; 4 hr; 37oC incubation
8. Loading buffer; 1 uL; 0.7X;
9. Incubation: 1hr @ RT in Dark.
10. Compounds; 23 nL; 0.59 nM to 92 uM
11. Controls; 23 nL; Astemizole 1.4 nM to 92 uM
12. Time; 10min; 37oC incubation
13. Read fluorescence Intensity on FDSS for 10 Sec with 1 sec interval
14. Reagent; 1 uL; stimulation buffer
15. Read fluorescence Intensity on FDSS for 2 min with 1 sec interval
16. Detection; Fluorescence Intensity; FDSS

Comment: Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

External ID: CHEMBL2426918

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426917

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426916

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426915

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL820514

Protocol: N/A

Comment: Compounds with activity <= 10uM or explicitly reported as active by ChEMBL are flagged as active in this PubChem assay presentation.

Journal: J. Med. Chem.
Year: 2003
Volume: 46
Issue: 8
First Page: 1478
Last Page: 1483
DOI: 10.1021/jm020427b

External ID: CHEMBL2426922

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: SCA2002

Protocol: HEK293 cultured cells will be dosed with titrated compounds and incubated for 72 hr at 37 degrees C. Toxicity will be assessed using the CellTiter Glo kit (Promega), which uses intracellular ATP concentration as a viability marker.

Comment: Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

External ID: CHEMBL2426921

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426920

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426919

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426910

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429625

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429624

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2429623

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426914

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426913

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426912

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426911

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426925

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426924

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL2426923

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2013
Volume: 76
Issue: 9
First Page: 1724
Last Page: 1730
DOI: 10.1021/np400433g

Target ChEMBL ID: CHEMBL383
ChEMBL Target Name: HL-60
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: SCA2001

Protocol: 4 ul of HEK293-ATXN2-FFLuc cell suspension (TBD cells/microL in phenol-red free DMEM) are dispensed into Greiner white solid bottom 1536-well assay plates, yielding a final well density of TBD cells/well. Plates are incubated for 2 hrs at 37 degrees C in 5% humidified CO2 to allow cell attachment. Compounds are then transferred via Kalypsys pin tool equipped with 1536-pin array (10nl slotted pins, V&P Scientific, San Diego, CA). In addition, duplicate serial dilutions of the control compounds ChemBridge 5553825, a previously identified ATXN2 expression inhibitor, and a known luciferase inhibitor, are pin-transferred. After incubation at 37 degrees C in 5% humidified CO2 for 24 hrs, CellTiter-Fluor reagent (1 ul) (Promega, Madison, WI) is then added and plates are incubated for 30 minutes, before being transferred to a ViewLux high-throughput CCD imager (PerkinElmer), wherein single end-point fluorescence measurements are acquired using the 405 nm excitation/525 nm emission filter set to assess cell viability. Next, Bright-Glo luciferase substrate detection reagent (3 ul) (Promega, Madison, WI) is added to each well and incubated for an additional 5 minutes. Luminescence is then measured on the ViewLux imager equipped with a clear emission filter using a 2 sec exposure.

Comment: Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

External ID: VIFG001

Protocol: 5 uL of 700 cells/well of A3G-YFP cells are dispensed into black, clear bottom plates. The plates are incubated overnight at 37 deg C. 23 nL of compounds are added by pintool. The plates are incubated overnight at 37 deg C and then read on the TTP Acumen (Ex 488/ Em 500-530).

Comment: Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

External ID: ERR513

Protocol: Tox21 Assay Protocol Summary:

The ERR cells were dispensed at 2,000 cells/5 ul/well in 1536-well white plates using a Multidrop dispenser. After the assay plates were incubated at a 37 C/5% CO2 incubator for 6 hours, 23 nL of compounds dissolved in DMSO, positive and negative controls or DMSO only was transferred to the assay plate by a pin tool. The plates were incubated at 37 C for 17.5 hours. 1 ul/well of CellTiter-Fluor reagent was added into the assay plates using a Flying Reagent Dispenser. After 30 min incubation at 37 C/5% CO2, the fluorescence intensity in the plates was measured using a ViewLux plate reader.

Comment: Disclaimer:

Although all reasonable efforts have been made to ensure the accuracy and reliability of the data, caution should be exercised when interpreting the results as artifacts are possible from nonspecific effects such as assay signal interference. The curve fitting and activity calls presented here are based on the NCATS analysis methods. Alternative analysis methods and interpretations of the data are available at EPA (http://actor.epa.gov) and NTP (http://tools.niehs.nih.gov/cebs3/ui/).

Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

External ID: CHEMBL960996

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 2009
Volume: 52
Issue: 7
First Page: 2067
Last Page: 2075
DOI: 10.1021/jm801605r

Target ChEMBL ID: CHEMBL1255151
ChEMBL Target Name: AmpC
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: ERR845

Protocol: Tox21 Assay Protocol Summary:

The ERR cells were dispensed at 2,000 cells/5 ul/well in 1536-well white plates using a Multidrop dispenser. After the assay plates were incubated at a 37 C/5% CO2 incubator for 6 hours, 23 nL of compounds dissolved in DMSO, positive and negative controls or DMSO only was transferred to the assay plate by a pin tool. The plates were incubated at 37 C for 18 hours. 4 ul/well of One-Glo reagent was added into the assay plates using a Flying Reagent Dispenser. After 30 min incubation at room temperature, the luminescence intensity in the plates was measured using a ViewLux plate reader.

Comment: Disclaimer:

Although all reasonable efforts have been made to ensure the accuracy and reliability of the data, caution should be exercised when interpreting the results as artifacts are possible from nonspecific effects such as assay signal interference. The curve fitting and activity calls presented here are based on the NCATS analysis methods. Alternative analysis methods and interpretations of the data are available at EPA (http://actor.epa.gov) and NTP (http://tools.niehs.nih.gov/cebs3/ui/).

Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.

External ID: CHEMBL960997

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 2009
Volume: 52
Issue: 7
First Page: 2067
Last Page: 2075
DOI: 10.1021/jm801605r

Target ChEMBL ID: CHEMBL1255151
ChEMBL Target Name: AmpC
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: CHEMBL960994

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 2009
Volume: 52
Issue: 7
First Page: 2067
Last Page: 2075
DOI: 10.1021/jm801605r

Target ChEMBL ID: CHEMBL1255151
ChEMBL Target Name: AmpC
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: 20160513eIF4E

Protocol: The cells carrying the bicitronic reporter gene (E8-Luc) were plated in 96-well plates at a density of 1.2x10(5)/well (100 ul DMEM) overnight. 0.5 ul of tested compounds (0.5 mM) or DMSO were added into the plates, and treated the cells for 16 h. The plates were then washed with 100 ul of PBS twice using Aquamax Plate Washer (Molecular Devices), and lysated in 60 ul of lysis buffer (1% Triton X-100, 25 mM Gly-Gly, pH 7.8, 15 mM MgSO4, and 4 mM EGTA) at room temperature for 20 min. 10 ul of lysates were then subjected to luciferase activity assays using Spectramax L (Molecular Devices) and the Promega Luciferase Substrate. The relative luciferase activities were then converted into logarithm values (binary logarithm, i.e., log2).

The cutoff value set for positives is 0.5849, which indicates that a compound can decrease the luciferase activity to at least 66.7% of the DMSO group.

Comment:

External ID: CHEMBL4390571

Protocol: N/A

Comment: Compounds with activity <= 10uM or explicitly reported as active by ChEMBL are flagged as active in this PubChem assay presentation.

Journal: J Nat Prod
Year: 2019
Volume: 82
Issue: 8
First Page: 2174
Last Page: 2180
DOI: 10.1021/acs.jnatprod.9b00182

Target ChEMBL ID: CHEMBL614421
ChEMBL Target Name: Micrococcus luteus
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL960995

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 2009
Volume: 52
Issue: 7
First Page: 2067
Last Page: 2075
DOI: 10.1021/jm801605r

Target ChEMBL ID: CHEMBL1255151
ChEMBL Target Name: AmpC
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

External ID: PGC981

Protocol: Tox21 Assay Protocol Summary:

The PGC/ERR cells were dispensed at 2,000 cells/5 ul/well in 1536-well white plates using a Multidrop dispenser. After the assay plates were incubated at a 37 C/5% CO2 incubator for 6 hours, 23 nL of compounds dissolved in DMSO, positive and negative controls or DMSO only was transferred to the assay plate by a pin tool. The plates were incubated at 37 C for 17.5 hours. 1 ul/well of CellTiter-Fluor reagent was added into the assay plates using a Flying Reagent Dispenser. After 30 min incubation at 37 C/5% CO2, the fluorescence intensity in the plates was measured using a ViewLux plate reader.

Comment: Disclaimer:

Although all reasonable efforts have been made to ensure the accuracy and reliability of the data, caution should be exercised when interpreting the results as artifacts are possible from nonspecific effects such as assay signal interference. The curve fitting and activity calls presented here are based on the NCATS analysis methods. Alternative analysis methods and interpretations of the data are available at EPA (http://actor.epa.gov) and NTP (http://tools.niehs.nih.gov/cebs3/ui/).

Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.