External ID: CHEMBL5134766

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

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: CHEMBL5134767

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134760

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134761

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134762

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134756

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134757

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134758

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134759

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134752

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134753

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134754

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134755

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: s-my-keats_OPM1-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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: CHEMBL5134748

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: s-my-OC1MY5-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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: CHEMBL5134749

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134750

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134751

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5514053

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2024
Volume: 268
First Page: 116273
Last Page: 116273
DOI: 10.1016/j.ejmech.2024.116273

Target ChEMBL ID: CHEMBL6006
ChEMBL Target Name: Serine/threonine-protein kinase ULK1
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: s-my-KMS_34-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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: CHEMBL5134744

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: s-my-keats_L363-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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: CHEMBL5134745

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: s-my-keats_OCIMY7-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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: CHEMBL5134746

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5134747

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5057014

Protocol: N/A

Comment: Journal: Bioorg Med Chem
Year: 2021
Volume: 49
First Page: 116456
Last Page: 116456
DOI: 10.1016/j.bmc.2021.116456

Target ChEMBL ID: CHEMBL614040
ChEMBL Target Name: J774.A1
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: s-my-MM1R-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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: CHEMBL5134740

Protocol: N/A

Comment: Journal: Eur J Med Chem
Year: 2022
Volume: 231
First Page: 114108
Last Page: 114108
DOI: 10.1016/j.ejmech.2022.114108

Target ChEMBL ID: CHEMBL354
ChEMBL Target Name: Escherichia coli
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

External ID: CHEMBL5057013

Protocol: N/A

Comment: Journal: Bioorg Med Chem
Year: 2021
Volume: 49
First Page: 116456
Last Page: 116456
DOI: 10.1016/j.bmc.2021.116456

Target ChEMBL ID: CHEMBL614040
ChEMBL Target Name: J774.A1
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: s-my-mm1s-m4-1

Protocol: SEQUENCE No. (e.g., 1, 2, 3, etc.); PARAMETER (e.g., Cells, Incubation, Reagent, etc.); VALUE and DESCRIPTION.
1; Cells; 5 uL; white, solid bottom 1536-well Greiner assay plate.
2; Compounds; 23 nL; Kalypsis pin tool (Wako USA) equipped with a 1536-well pin head to transfer to the assay plates.
3; Incubation; 48 hours; 37C incubation.
4; Reagent;5uL; CellTiter Glo (Promega).
5; Detector; 425V; Luminescence; Viewlux PerkinElmer.

NOTES (numbers refer to Sequence numbers above)
1. The cell-based screening methods employed in this study were like those previously published [1]. Briefly, the corresponding myeloma cell lines were screened in 1,536-well plates with 500 cells, 5-uL per well for inhibition of cell viability (assessed by measuring ATP levels with CellTiterGlo).
2. NCATS MIPE 4.0 library of approved and investigational drugs were added using the Kalypsis pin tool. Wells treated with DMSO only for 100% viability and media only for 0% viability were used as controls.
3. The assay plates were covered and incubated for 48 hours in a 37C, 5% CO2 incubator with controlled humidity.
4. After 48 hours, CellTilter-GloTM luminescent substrate mix (Promega) reagent was added to the assay plate and plates we incubated for 5 min at room temperature.
5. Signal was measured as median relative luminescence units on a ViewLux (PerkinElmer) multiplate reader.

Reference:
[1] Heske CM, Davis MI, Baumgart JT, Wilson K, Gormally MV, Chen L, Zhang X, Ceribelli M, Duveau DY, Guha R, Ferrer M, Arnaldez FI, Ji J, Tran HL, Zhang Y, Mendoza A, Helman LJ, Thomas CJ. Matrix Screen Identifies Synergistic Combination of PARP Inhibitors and Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibitors in Ewing Sarcoma. Clin Cancer Res. 2017 Dec 1;23(23):7301-7311. doi: 10.1158/1078-0432.CCR-17-1121. Epub 2017 Sep 12. PMID: 28899971; PMCID: PMC6636827.

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 cytotoxic compounds are ranked higher than compounds that showed no activity.
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.