HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262461

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.25	ug.mL-1
MIC	=	32	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	0.125	ug.mL-1
MIC	=	0.028	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262460

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.25	ug.mL-1
MIC	=	32	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	0.06	ug.mL-1
MIC	=	0.028	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262459

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.25	ug.mL-1
MIC	=	32	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	0.06	ug.mL-1
MIC	=	0.028	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262458

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.5	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.007	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 8

External ID: USP8 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP8: Primary Screen
1.#Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2.#Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3.#Reagent, 2.5 uL of USP8 in assay buffer (final concentration of 1 nM) to columns 1-46
4.#Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 50 nM) to all wells
5.#Time, Incubate at room temperature for 30 minutes
6.#Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP8 (USP8_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

USP8-Efficacy
46.639
-12.507
31.998
5.438
-6.685
15.21
-15.155
14.048
-4.808
14.043
13.854
55.661
-5.386
27.936
7.845
12.88
0.228
3.333
-3.366
0.283

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262457

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.5	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	8	ug.mL-1
MIC	=	0.007	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262456

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.5	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.007	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 17 like family member 5

External ID: USP17 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP17: Primary Screen
1. Compound, 50 nL of screening compounds (final concentration of 50 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP17 in assay buffer (final concentration of 1 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
5. Time, Incubate at room temperature for 3 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP17 (USP17_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

USP17-Efficacy
-11.703
-22.774
-17.896
1.991
13.524
0.0347
-9.782
55.109
-7.352
14.263
28.858
-17.021
12.515
47.032
7.524
3.661
-9.31
-4.13
17.206
2.868

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262455

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.5	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	8	ug.mL-1
MIC	=	0.007	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Cryptococcus gattii

External ID: CHEMBL1261944

Protocol: N/A

Comment: Journal: Antimicrob. Agents Chemother.
Year: 2010
Volume: 54
Issue: 6
First Page: 2303
Last Page: 2311
DOI: 10.1128/aac.00153-10

Target ChEMBL ID: CHEMBL615035
ChEMBL Target Name: Cryptococcus bacillisporus
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Activity Comment
Activity	Not Active

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 7

External ID: USP7 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP7: Primary Screen
1. Reagent, 2.5 uL of USP7 in assay buffer (final concentration of 4 nM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076)
2. Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 (columns 45-46: DMSO control)
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Time, Incubate at room temperature for 60 minutes
5. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
6. Time, Incubate at room temperature for 10 minutes
7. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP7 (USP7_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

USP7_EFFICACY
-0.801
-6.934
-7.543
2.861
2.363
-17.749
-0.834
-2.389
0.879
0.233
2.8
-8.773
-0.815
-0.271
7.414
-5.634
0.496
3.06
-13.572
5.477

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Saccharomyces cerevisiae

External ID: CHEMBL1262454

Protocol: N/A

Comment: Journal: Antimicrob Agents Chemother
Year: 2008
Volume: 52
Issue: 4
First Page: 1325
Last Page: 1329
DOI: 10.1128/aac.01393-07

Target ChEMBL ID: CHEMBL361
ChEMBL Target Name: Saccharomyces cerevisiae
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	0.5	ug.mL-1
MIC	=	16	ug.mL-1
MIC	=	0.025	ug.mL-1
MIC	=	8	ug.mL-1
MIC	=	0.007	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Cryptococcus gattii

External ID: CHEMBL1261943

Protocol: N/A

Comment: Journal: Antimicrob. Agents Chemother.
Year: 2010
Volume: 54
Issue: 6
First Page: 2303
Last Page: 2311
DOI: 10.1128/aac.00153-10

Target ChEMBL ID: CHEMBL615035
ChEMBL Target Name: Cryptococcus bacillisporus
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Activity Comment
Activity	Not Active

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Hypoxia-inducible factor 1-alpha

External ID: CHEMBL3854217

Protocol: N/A

Comment: Journal: J Nat Prod
Year: 2016
Volume: 79
Issue: 5
First Page: 1267
Last Page: 1275
DOI: 10.1021/acs.jnatprod.5b00846

Target ChEMBL ID: CHEMBL4261
ChEMBL Target Name: Hypoxia-inducible factor 1 alpha
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

Standard Type	Standard Units
Inhibition	%

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Hypoxia-inducible factor 1-alpha

External ID: CHEMBL3854213

Protocol: N/A

Comment: Journal: J Nat Prod
Year: 2016
Volume: 79
Issue: 5
First Page: 1267
Last Page: 1275
DOI: 10.1021/acs.jnatprod.5b00846

Target ChEMBL ID: CHEMBL4261
ChEMBL Target Name: Hypoxia-inducible factor 1 alpha
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

Standard Type	Standard Units	Activity Comment
Inhibition	%	Active

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Hypoxia-inducible factor 1-alpha

External ID: CHEMBL3854214

Protocol: N/A

Comment: Journal: J Nat Prod
Year: 2016
Volume: 79
Issue: 5
First Page: 1267
Last Page: 1275
DOI: 10.1021/acs.jnatprod.5b00846

Target ChEMBL ID: CHEMBL4261
ChEMBL Target Name: Hypoxia-inducible factor 1 alpha
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

Standard Type	Standard Units	Activity Comment
Inhibition	%	Active

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Hypoxia-inducible factor 1-alpha

External ID: CHEMBL3854215

Protocol: N/A

Comment: Journal: J Nat Prod
Year: 2016
Volume: 79
Issue: 5
First Page: 1267
Last Page: 1275
DOI: 10.1021/acs.jnatprod.5b00846

Target ChEMBL ID: CHEMBL4261
ChEMBL Target Name: Hypoxia-inducible factor 1 alpha
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

Standard Type	Standard Units
Inhibition	%

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Mus musculus

External ID: CHEMBL732279

Protocol: N/A

Comment: Journal: Bioorg. Med. Chem. Lett.
Year: 1997
Volume: 7
Issue: 20
First Page: 2645
Last Page: 2650
DOI: 10.1016/S0960-894X(97)10052-X

Target ChEMBL ID: CHEMBL375
ChEMBL Target Name: Mus musculus
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
Activity	=	59	%

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Hypoxia-inducible factor 1-alpha

External ID: CHEMBL3854216

Protocol: N/A

Comment: Journal: J Nat Prod
Year: 2016
Volume: 79
Issue: 5
First Page: 1267
Last Page: 1275
DOI: 10.1021/acs.jnatprod.5b00846

Target ChEMBL ID: CHEMBL4261
ChEMBL Target Name: Hypoxia-inducible factor 1 alpha
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: D - Direct protein target assigned
Confidence: Direct single protein target assigned

Standard Type	Standard Units
Inhibition	%

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Creatine kinase M-type

External ID: CHEMBL664050

Protocol: N/A

Comment: Journal: Bioorg. Med. Chem. Lett.
Year: 1997
Volume: 7
Issue: 20
First Page: 2645
Last Page: 2650
DOI: 10.1016/S0960-894X(97)10052-X

Target ChEMBL ID: CHEMBL2656
ChEMBL Target Name: Creatine kinase M
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

Standard Type	Standard Relation	Standard Value	Standard Units
Activity	=	32	%

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Candida albicans

External ID: CHEMBL1000905

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: 2005
Volume: 68
Issue: 8
First Page: 1300
Last Page: 1302
DOI: 10.1021/np050141k

Target ChEMBL ID: CHEMBL366
ChEMBL Target Name: Candida albicans
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
0.028	IC50	=	28	nM
0.083	IC50	=	83	nM
31	IC50	>	31000	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：CCRF-CEM

External ID: CHEMBL1000906

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: 2005
Volume: 68
Issue: 8
First Page: 1300
Last Page: 1302
DOI: 10.1021/np050141k

Target ChEMBL ID: CHEMBL382
ChEMBL Target Name: CCRF-CEM
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
0.58	IC50	=	580	nM
4.2	IC50	=	4200	nM
31	IC50	>	31000	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：NCGC 靶标：gene 4 small orf - Marburg virus

External ID: VSVM-OFFLINE

Protocol: Two uL of HEK293 cell suspension are dispensed at 1000 cells/well into solid white 1536-well plates (Grenier) using a Multidrop Combi (Thermo Scientific). After addition of 23 nL compound by a pin tool (Kalypsys), the plate is incubated 1 h at 37 degrees C and then 3 uL of virus 1:100 dilution VSV-MARV is added. After 28 hr, 4 uL of assay reagent is added and the plates are read using a ViewLux (Perkin Elmer). Assays are performed in sub-saturating amounts of virus (MOI <0.5), therefore luciferase signals reflect the amount (titer) of virus able to infect the cells in presence of the compound.

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.

Phenotype	Potency	Efficacy	Activity_Score	Curve_Description	Fit_LogAC50	Fit_HillSlope	Fit_R2	Fit_InfiniteActivity	Fit_ZeroActivity	Fit_CurveClass	Excluded_Points	Max_Response	Activity at 0.076 uM	Activity at 0.631 uM	Activity at 1.728 uM	Activity at 8.589 uM	Activity at 49.20 uM	Compound QC
Inactive			0			4.9549	0.4153	-14.5146	1	4	0 0 0 0 1	4.685	-8.5335	0.7807	11.3474	-12.5122	4.685	QC'd by "Microsource"
Activator	12.5893	29.2705	0	Single point of activity	-4.9	4.9549	0.9612	31	1.7295	3	0 0 0 0 0	30.9001	-1.4754	1.0166	5.6243	6.6083	30.9001	QC'd by "Microsource"
Activator	8.9125	65.9699	0	Partial curve; partial efficacy	-5.05	0.6	0.9478	60.7629	-5.207	2.2	0 0 0 0 0	46.0767	-2.3225	0.7459	18.7564	22.5194	46.0767	QC'd by "Microsource"
Inactive			0			4.9549	0.3508	-5.4241	12	4	0 0 0 0 0	2.824	6.9985	-6.3081	-17.4367	-1.0266	2.824	QC'd by "Microsource"
Inactive			0			4.095	0.8518	-20.6942	-4.8432	4	1 0 0 0 0	-16.9639	-25.916	-5.286	-19.6983	-24.7451	-16.9639	QC'd by "Microsource"
Inactive			0			4.9549	0.8518	-16.2964	1.5	4	0 0 0 0 1	4.1206	1.8961	-0.6171	-21.497	-11.4248	4.1206	QC'd by "Microsource"
Activator	2.2387	83.6659	0	Partial curve; partial efficacy; poor fit	-5.65	0.5	0.8806	54.0974	-29.5685	2.4	0 0 0 0 0	44.3794	-19.3312	-4.1329	18.8496	13.5792	44.3794	QC'd by "Microsource"
Inhibitor	12.5893	81.1747	42	Partial curve; high efficacy	-4.9	4.095	0.9407	-88.2352	-7.0605	-2.1	0 0 0 0 0	-88.059	-2.1141	2.5457	-21.0606	-24.383	-88.059	QC'd by "Microsource"
Inactive			0			0.8	0.9754	0.5	18	4	0 0 0 0 0	0.1686	15.5939	9.5523	5.2389	3.3969	0.1686	QC'd by "Microsource"
Inactive			0			4.9549	0.516	14	-11.1922	4	0 0 0 0 0	8.9233	2.0681	-9.5798	-21.4101	-15.067	8.9233	QC'd by "Microsource"
Inhibitor	10	111.4578	10	Partial curve; high efficacy; poor fit	-5	4.095	0.8557	-88.0204	23.4373	-2.3	0 0 0 0 0	-86.963	0.1371	16.1149	55.0816	-24.8092	-86.963	QC'd by "Microsource"
Inhibitor	12.5893	111.8609	10	Single point of activity	-4.9	1.3987	0.9539	-66.6934	45.1675	-3	0 0 0 0 0	-51.9056	34.8895	58.2591	32.624	3.3045	-51.9056	QC'd by "Microsource"
Inactive			0							4		9.4501	23.4721	50.1914	-0.8776	27.4967	9.4501	QC'd by "Microsource"
Inactive			0			3.6272	0.9908	-9.6077	22.5	4	0 0 0 0 0	-9.2564	21.9907	20.6042	24.2232	8.6092	-9.2564	QC'd by "Microsource"
Activator			0	Single point of activity		4.9549	0.3547	25.2512	5	3	0 0 0 0 1	15.4142	5.7176	6.5342	43.2316	7.0919	15.4142	QC'd by "Microsource"
Inactive			0			4.9549	0.6074	7	18.5	4	0 0 0 0 0	11.6515	12.5897	21.5938	21.2209	4.3118	11.6515	QC'd by "Microsource"
Activator	2.2387	52.7895	0	Partial curve; partial efficacy	-5.65	2.2526	0.9613	44.2358	-8.5537	2.2	0 0 0 0 1	-9.4264	-2.4981	-13.182	11.9591	42.1127	-9.4264	QC'd by "Microsource"
Inactive			0			4.9549	0.942	28	1.1909	4	0 0 0 0 1	1.5316	4.6729	1.2372	-2.341	24.4478	1.5316	QC'd by "Microsource"
Inhibitor	35.4813	55.7622	10	Single point of activity	-4.45	4.9549	0.7876	-42.7915	12.9707	-3	0 0 0 0 0	-30.6839	14.5151	3.8713	4.7738	29.0235	-30.6839	QC'd by "Microsource"
Inhibitor	35.4813	49.2317	10	Single point of activity	-4.45	4.4495	0.7066	-51.9324	-2.7008	-3	0 0 0 0 0	-40.193	-17.9891	11.8148	-5.5992	0.7455	-40.193	QC'd by "Microsource"

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：NCGC 靶标：N/A

External ID: VSVL-OFFLINE

Protocol: For screening, 1000 cells in 2 ul/well were dispensed into white solid 1536-well plates (Greiner) using a solenoid-based dispenser. Following transfer of 23nl compound or DMSO vehicle by a pin tool, 3 ul/well of VSV-LV was added. The plates were centrifuged 1 min at 1000 RPM and then incubated 16 hr at 37 C and 5% CO2. After addition of 4 ul/well SteadyLite (PerkinElmer) detection reagent, the plates were incubated 10 min at ambient temperature and luminescence was measured on a ViewLux (Perkin Elmer) plate reader.

Keywords: NIH Roadmap, MLPCN, MLI, MLSMR, qHTS, NCGC, Lassa virus, luciferase, cell assay, infection

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.

Phenotype	Potency	Efficacy	Activity_Score	Curve_Description	Fit_LogAC50	Fit_HillSlope	Fit_R2	Fit_InfiniteActivity	Fit_ZeroActivity	Fit_CurveClass	Excluded_Points	Max_Response	Activity at 0.631 uM	Activity at 1.728 uM	Activity at 3.886 uM	Activity at 8.587 uM	Activity at 17.80 uM	Activity at 49.20 uM	Compound QC
Inhibitor	25.1189	46.3945	20	Partial curve; partial efficacy	-4.6	2.2526	0.9287	-51.2557	-4.8612	-2.2	0 0 0 0 0	-41.1028		-5.7868	-8.9838	-1.9677	-26.4188	-41.1028	QC'd by "BIOMOL"
Inactive			0			3.5117	0.9435	10	-4.567	4	0 0 0 0 1	-5.6299	-3.6518	-3.5753	-4.2225	6.795	-5.6299		QC'd by "Prestwick Chemical; Inc."
Inactive			0			3.0654	0.9376	11	-2.7695	4	0 0 0 0 1	-2.8045	-1.4193	-3.1412	-1.9048	7.584	-2.8045		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.7625	1	-16.7	4	0 0 0 0	-0.288	-12.25	5.019	-2.495		-0.288		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.8335	4.5	-7.0946	4	0 0 0 0	3.0138	-4.2455	6.6652	3.4342		3.0138		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.993	0.5	-16.2895	4	0 0 0 0 1	-17.2531	-11.9079	0.4577	0.7459	0.1472	-17.2531		QC'd by "Prestwick Chemical; Inc."
Inactive			0			3.0654	0.989	-13.9862	-8.1964	4	0 0 0 0	-13.7385	-8.6044	-8.0804	-9.4725		-13.7385		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.8435	-11.2843	4	4	0 0 0 0 0	-11.9036	3.4878	5.2393	0.5291	5.9839	-11.9036		QC'd by "Prestwick Chemical; Inc."
Inactive			0			0.8	0.7241	0.6	-14.6999	4	0 0 0 0 1	-12.1307	-11.4165	-6.1179	-8.0206	-2.8333	-12.1307		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.8652	1	-19.7041	4	0 0 0 0 0	-4.2947	-18.0867	-15.9371	4.8395	2.1157	-4.2947		QC'd by "Prestwick Chemical; Inc."
Inhibitor	14.1254	42.1465	10	Single point of activity	-4.85	4.9549	0.8997	-40.1465	2	-3	0 0 0 0 0	-41.8192	-2.393	5.1804	-2.5248	4.6867	-41.8192		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.6566	-10.2454	0	4	0 0 0 0 0	-10.2045	-4.5492	-0.0989	4.6103	-5.6017	-10.2045		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.5279	3	-11.3522	4	0 0 0 0	1.078	-15.7102	-2.9932	-14.9547		1.078		QC'd by "Prestwick Chemical; Inc."
Inactive			0			1.6924	0.9123	0.5	-10.147	4	0 0 0 0	0.4297	-9.2892	-3.9336	-3.1566		0.4297		QC'd by "Prestwick Chemical; Inc."
Inactive			0			2.4064	0.7886	8	-4.3029	4	0 0 0 0 0	5.7256	-1.9822	-4.4191	-3.8175	8.0142	5.7256		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.997	5.5	-12.076	4	0 0 0 1	-6.5693	-12.5633	-7.4947	4.9219		-6.5693		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.9232	-18.4489	-2.2837	4	0 0 0 0	-16.2075	-1.3456	-4.9534	-0.2364		-16.2075		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.9481	2	-17.5643	4	0 0 0 0 1	-13.5121	-12.9703	3.6247	1.4507	0.7522	-13.5121		QC'd by "Prestwick Chemical; Inc."
Inactive			0			4.9549	0.5725	-10.2193	7.5	4	0 0 0 0 0	-12.7555	5.891	7.5423	-18.5161	0.6587	-12.7555		QC'd by "Prestwick Chemical; Inc."
Inhibitor	31.6228	100.7069	40	Partial curve; high efficacy	-4.5	3.5722	0.9933	-104.0783	-3.3715	-2.1	0 0 0 0 0	-83.3961		0.3188	-7.6251	-4.9588	-20.5702	-83.3961	QC'd by "BIOMOL"

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Histone-lysine N-methyltransferase EHMT2

External ID: CHEMBL1953573

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: 2012
Volume: 75
Issue: 1
First Page: 111
Last Page: 114
DOI: 10.1021/np200740e

Target ChEMBL ID: CHEMBL6032
ChEMBL Target Name: Histone-lysine N-methyltransferase, H3 lysine-9 specific 3
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
2.1	IC50	=	2100	nM
100	IC50	>	100000	nM
2.6	IC50	=	2600	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：P388

External ID: CHEMBL1953572

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: 2012
Volume: 75
Issue: 1
First Page: 111
Last Page: 114
DOI: 10.1021/np200740e

Target ChEMBL ID: CHEMBL389
ChEMBL Target Name: P388
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
0.02	IC50	=	20	nM
3.4	IC50	=	3400	nM
0.056	IC50	=	56	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Geranylgeranyl transferase type-1 subunit beta

External ID: CHEMBL686125

Protocol: N/A

Comment: Journal: Bioorg. Med. Chem. Lett.
Year: 2003
Volume: 13
Issue: 21
First Page: 3661
Last Page: 3663
DOI: 10.1016/j.bmcl.2003.08.022

Target ChEMBL ID: CHEMBL2095164
ChEMBL Target Name: Geranylgeranyl transferase type I
ChEMBL Target Type: PROTEIN COMPLEX - Target is a defined protein complex, consisting of multiple subunits
Relationship Type: D - Direct protein target assigned
Confidence: Direct protein complex subunits assigned

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
100	IC50	>	100000	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Histone-lysine N-methyltransferase SETD7

External ID: CHEMBL1953574

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2012
Volume: 75
Issue: 1
First Page: 111
Last Page: 114
DOI: 10.1021/np200740e

Target ChEMBL ID: CHEMBL5523
ChEMBL Target Name: Histone-lysine N-methyltransferase SETD7
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

Standard Type	Standard Units	Activity Comment
Inhibition	%	Not Active
Inhibition	%	Not Active
Inhibition	%	Not Active
Inhibition	%	Not Active

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：N/A

External ID: CHEMBL699669

Protocol: N/A

Comment: Journal: J. Med. Chem.
Year: 1997
Volume: 40
Issue: 19
First Page: 2971
Last Page: 2990
DOI: 10.1021/jm970226l

Standard Type	Standard Relation	Standard Value	Standard Units
Concentration	=	6.3	mg kg-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：NCGC 靶标：DNA polymerase kappa [Homo sapiens]

External ID: PolK100

Protocol: Three microliters of reagents (buffer in column 3 and 4 as negative control and 10 nM Pol kappa in columns 1, 2, and 5-48) were dispensed into a 1536-well black solid-bottom plate. Compounds (23 nL) were transferred via Kalypsys pin tool equipped with 1536-pin array. The plates were then incubated for 15 min at room temperature, and 1 uL substrate (50 nM final concentration) were then added to start the reaction and kinetically read twice at 0 min and 10 min on the Viewlux reader

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.

Phenotype	Fit_CurveClass	Excluded_Points	Max_Response	Activity at 0.477 uM	Activity at 2.393 uM	Activity at 11.99 uM	Activity at 60.11 uM	Activity at 107.2 uM	Compound QC
Inactive	4	0 0 0 0 0	1.4694	-3.5669	-6.235	2.8586	1.8042	1.4694	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	-4.2631	8.2218	8.0811	10.2927	-3.9947	-4.2631	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	6.0369	0.3398	-2.1048	-8.1695	-3.6822	6.0369	QC'd by "Chem Div"
Inactive	4		-2.0565	1.7294	-3.5894	-1.2575	-0.5402	-2.0565	QC'd by "Chem Div"
Inactive	4	0 0 0 0 1	2.3149	1.0048	4.6369	-1.9963	-3.3543	2.3149	QC'd by "Chem Div"
Inactive	4		7.2748	7.1515	6.1372	1.5197	5.2332	7.2748	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	1.006	-3.3873	-7.786	-9.3037	-9.1761	1.006	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	-0.0368	-9.4458	-10.5155	-9.0065	-12.9141	-0.0368	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	2.6	-7.8084	-12.3007	-2.0954	-6.6887	2.6	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	-11.4867	-18.9051	-17.4955	-19.0735	-9.6682	-11.4867	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	-7.5605	-17.2173	-11.0038	-16.5656	-22.4025	-7.5605	QC'd by "Chem Div"
Inactive	4		-7.5451	-1.1939	-1.3084	-5.8268	-5.3206	-7.5451	QC'd by "Chem Div"
Inactive	4	0 0 0 0 1	-5.5852	-4.3753	-1.0046	-3.1641	-10.1524	-5.5852	QC'd by "Chem Div"
Inactive	4	0 0 0 0 0	1.1172	-6.0391	7.0118	9.0446	1.6533	1.1172	QC'd by "Chem Div"
Inactive	4		2.3359	1.2518	1.6626	-0.9325	-0.9194	2.3359	QC'd by "Chem Div"
Inactive	4	0 0 0 0	-19.5354	0.3984	-4.1147	2.1883	-19.5354		QC'd by "Chem Div"
Inactive	4		-5.6552	-4.6769	-1.9378	-0.5867	-3.224	-5.6552	QC'd by "Chem Div"
Inactive	4		-11.3738	-10.4148	-13.8912	-10.4252	-7.8961	-11.3738	QC'd by "Chem Div"
Inactive	4		-6.1571	-8.7102	-2.9113	-5.2229	-3.4369	-6.1571	QC'd by "Chem Div"
Inactive	4	0 0 0 0 1	-7.3803	-8.8177	-11.1654	-6.5301	-15.9483	-7.3803	QC'd by "Chem Div"

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Candida albicans

External ID: CHEMBL1921020

Protocol: N/A

Comment: Journal: J. Nat. Prod.
Year: 2011
Volume: 74
Issue: 8
First Page: 1721
Last Page: 1730
DOI: 10.1021/np2001573

Target ChEMBL ID: CHEMBL366
ChEMBL Target Name: Candida albicans
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Standard Relation	Standard Value	Standard Units
MIC	=	4	ug.mL-1

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 30

External ID: USP30 FAST DUB HTS Confirmation

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP30: Confirmation Screen
1. Compounds in 4-point log dose response were transferred from source plates to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) starting at an initial concentration of 50 uM (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of USP30 in assay buffer (final concentration of 1 nM) to columns 1-46
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 12 nM) to all wells
5. Time, Incubate at room temperature for 2 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Each compound was tested in triplicate in the dose-response confirmation assay. Normalization of raw data to positive and negative control wells, systematic pattern models, and curve fitting were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018). For measuring compound potency, a four-parameter sigmoid Hill curve model was fitted to the data (Gubler et al., 2018) and the absolute IC50 was used for potency measurements, that is the concentration of compound where the data showed 50% inhibition. If no data points achieved 50% inhibition, then the absolute IC50 was reported as >50 uM (the highest tested concentration). Maximal efficacy of each compound was reported as a percent inhibition based on normalization to plate controls.

Primary screen hits were confirmed as active if they demonstrated greater than or equal to 30% inhibition of USP30 (USP30_EFFICACY >/= 30). Active compounds were given a score based on potency.

Qualifier	USP30_Potency	USP30_Efficacy
=	43.119076	61.498283
>	50	40.273956
=	7.1207333	56.356796
=	22.86257	77.42735
>	50	-21.63186
>	50	-346.2283
=	9.6289673	77.21069
>	50	19.738224
>	50	-310.2163
>	50	27.170631
>	50	8.648887
=	30.767855	62.49963
>	50	-12.809548
>	50	-0.63047993
>	50	46.852955
>	50	-124.78407
>	50	40.666794
>	50	-19.45689
>	50	17.258204
>	50	30.849586

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 17 like family member 5

External ID: USP17 FAST DUB HTS Confirmation

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP17: Confirmation Screen
1. Compounds in 4-point, log dose response were transferred from source plates to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) starting at an initial concentration of 50 uM (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of USP17 in assay buffer (final concentration of 1 nM) to columns 1-46
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
5. Time, Incubate at room temperature for 3 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Each compound was tested in triplicate in the dose-response confirmation assay. Normalization of raw data to positive and negative control wells, systematic pattern models, and curve fitting were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018). For measuring compound potency, a four-parameter sigmoid Hill curve model was fitted to the data (Gubler et al., 2018) and the absolute IC50 was used for potency measurements, that is the concentration of compound where the data showed 50% inhibition. If no data points achieved 50% inhibition, then the absolute IC50 was reported as >50 uM (the highest tested concentration). Maximal efficacy of each compound was reported as a percent inhibition based on normalization to plate controls.

Primary screen hits were confirmed as active if they demonstrated greater than or equal to 30% inhibition of USP17 (USP17_EFFICACY >/= 30). Active compounds were given a score based on potency.

Qualifier	USP17_Potency	USP17_Efficacy
=	22.940981	67.466415
>	50	-52.472378
>	50	-105.95489
>	50	-8.899009
>	50	33.32131
>	50	-117.31118
>	50	-146.12769
=	5.8091812	64.42049
>	50	43.99427
>	50	-0.648086
=	20.82271	87.292534
=	25.277653	68.52241
=	43.723148	54.9599
>	50	2.750618
>	50	-50.683548
=	4.6738453	57.335205
>	50	20.037851
>	50	-159.9229
>	50	42.911022
=	22.926788	69.59911

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 28

External ID: USP28 FAST DUB HTS Confirmation

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP28: Confirmation Screen
1. Compounds in 4-point, log dose response were transferred from source plates to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) starting at an initial concentration of 50 uM (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of USP28 in assay buffer (final concentration of 0.5 nM) to columns 1-46
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 12 nM) to all wells
5. Centrifuge plates (5s, 350 rpm)
6. Time, Incubate at room temperature for 30 minutes
7. Detection, Fluorescence, PheraStar, FITC Module

Comment: Each compound was tested in triplicate in the dose-response confirmation assay. Normalization of raw data to positive and negative control wells, systematic pattern models, and curve fitting were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018). For measuring compound potency, a four-parameter sigmoid Hill curve model was fitted to the data (Gubler et al., 2018) and the absolute IC50 was used for potency measurements, that is the concentration of compound where the data showed 50% inhibition. If no data points achieved 50% inhibition, then the absolute IC50 was reported as >50 uM (the highest tested concentration). Maximal efficacy of each compound was reported as a percent inhibition based on normalization to plate controls.

Primary screen hits were confirmed as active if they demonstrated greater than or equal to 30% inhibition of USP28 (USP28_EFFICACY >/= 30). Active compounds were given a score based on potency.

Qualifier	USP28_Potency	USP28_Efficacy
=	33.249394	69.82961
=	19.943533	75.29613
=	12.391049	83.84166
=	23.614925	69.484474
>	50	-0.9354495
>	50	-70.89557
=	2.1291766	91.56192
=	12.462843	56.66996
>	50	-138.24467
=	17.432396	71.274475
=	13.053516	79.13997
=	30.557766	58.27259
=	3.8206532	88.29279
=	37.736027	61.918526
=	40.989094	56.682365
>	50	-0.38224816
=	40.782974	53.573406
>	50	28.136835
>	50	13.498848
=	12.610698	83.19416

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：OTU deubiquitinase 3

External ID: OTUD3 FAST DUB HTS Confirmation

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of OTUD3: Confirmation Screen
1. Compounds in 4-point, log dose response were transferred from source plates to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) starting at an initial concentration of 50 uM (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of OTUD3 in assay buffer (final concentration of 12 nM) to columns 1-46
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
5. Time, Incubate at room temperature for 2 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Each compound was tested in triplicate in the dose-response confirmation assay. Normalization of raw data to positive and negative control wells, systematic pattern models, and curve fitting were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018). For measuring compound potency, a four-parameter sigmoid Hill curve model was fitted to the data (Gubler et al., 2018) and the absolute IC50 was used for potency measurements, that is the concentration of compound where the data showed 50% inhibition. If no data points achieved 50% inhibition, then the absolute IC50 was reported as >50 uM (the highest tested concentration). Maximal efficacy of each compound was reported as a percent inhibition based on normalization to plate controls.

Primary screen hits were confirmed as active if they demonstrated greater than or equal to 30% inhibition of OTUD3 (OTUD3_EFFICACY >/= 30). Active compounds were given a score based on potency.

Qualifier	OTUD3_Potency	OTUD3_Efficacy
=	25.735994	75.8494
>	50	45.57201
=	25.651028	66.55539
>	50	14.195643
>	50	3.6956549
>	50	-191.96823
>	50	47.6023
>	50	3.9394982
>	50	-181.83427
>	50	4.321017
>	50	-12.092948
>	50	29.372139
>	50	-11.334712
>	50	27.327524
>	50	20.6282
>	50	-82.91289
>	50	16.600174
>	50	-10.991076
>	50	0.2047818
>	50	20.636448

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 28

External ID: USP28 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP28: Primary Screen
1. Compound, 25 nL of screening compounds (final concentration of 25 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP28 in assay buffer (final concentration of 0.5 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 12 nM) to all wells
5. Time, Incubate at room temperature for 30 minutes
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP28 (USP28_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

USP28-Efficacy
0.858
-79.548
-6.397
12.687
1.953
1.774
6.852
4.69
11.944
-0.62
6.642
-4.813
10.139
-3.985
6.898
5.589
87.267
-2.429
-1.878
1.221

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 10

External ID: USP10 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP10: Primary Screen
1. Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP10 in assay buffer (final concentration of 15 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 15 nM) to all wells
5. Time, Incubate at room temperature for 60 minutes
6. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP10 (USP10_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

USP10-Efficacy
3.101
1.907
-3.869
-7.539
0.549
-1.651
0.901
0.0502
0.956
15.834
-0.00175
-4.55
-1.07
1.85
2.992
9.647
0.745
-1.799
3.055
-0.917

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin C-terminal hydrolase L1

External ID: UCHL1 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of UCHL1: Primary Screen
1. Reagent, 2.5 uL of UCHL1 in assay buffer (final concentration of 1 nM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076)
2. Compound, 20 nL of screening compounds (final concentration of 20 uM) to columns 1-46 (columns 45-46: DMSO control)
3. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
4. Time, Incubate at room temperature for 60 minutes
5. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 7 nM) to all wells
6. Time, Incubate at room temperature for 10 minutes
7. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of UCHL1 (UCHL1_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

UCHL1-Efficacy
-6.982
0.0218
-4.751
4.329
-3.437
4.201
2.693
2.036
0.352
0.0185
-3.604
0.268
9.86
0.843
3.782
-17.728
4.105
11.718
6.238
-0.64

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：OTU deubiquitinase 3

External ID: OTUD3 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of OTUD3: Primary Screen
1. Compound, 25 nL of screening compounds (final concentration of 25 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of OTUD3 in assay buffer (final concentration of 12 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 40 nM) to all wells
5. Time, Incubate at room temperature for 2 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of OTUD3 (OTUD3_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

OTUD3-Efficacy
-7.265
7.071
-1.861
-3.016
-10.763
10.254
-10.529
4.602
-4.313
-5.645
0.343
8.993
-2.623
-1.623
-18.682
11.86
40.974
6.117
17.524
-56.479

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：24642 靶标：ubiquitin specific peptidase 30

External ID: USP30 FAST DUB HTS Primary

Protocol: Deubiquitinases were purified from E. Coli. Ub-Rho substrate was purchased from Boston BioChem (Cat #: U-555). Assay buffer consisted of 50 mM Tris pH 8, 50 mM NaCl, 0.002% Tween20, 5 mM DTT in MilliQ water. Stop buffer consisted of assay buffer + 0.2% trifluoroacetic acid.

Protocol for Inhibitors of USP30: Primary Screen
1. Compound, 25 nL of screening compounds (final concentration of 25 uM) to columns 1-46 of a Greiner 1536 well Black, flat bottom, polystyrene microplate (Griener 782076) (columns 45-46: DMSO control)
2. Reagent, 2.5 uL of assay buffer to columns 47-48 on all plates (No enzyme control)
3. Reagent, 2.5 uL of USP30 in assay buffer (final concentration of 1 nM) to columns 1-46
4. Reagent, 2.5 uL of Ub-Rho in assay buffer (final concentration of 12 nM) to all wells
5. Time, Incubate at room temperature for 2 hours
6. Reagent, 2.5 uL of stop buffer to all wells
7. Centrifuge plates (5s, 350 rpm)
8. Detection, Fluorescence, PheraStar, FITC Module

Comment: Normalization of raw data to positive and negative control wells were applied to the data using Helios, a high-throughput screening data analysis program developed at Novartis.(Gubler et al., 2018)

Compounds demonstrating greater than or equal to 30% inhibition of USP30 (USP30_EFFICACY >/= 30) were called as active, compounds demonstrating activity below this threshold were called as inactive.

USP30-Efficacy
34.821
21.39
49.071
-39.208
23.815
-0.611
-9.086
-8.452
-5.803
-4.031
-6.381
-5.714
0
-29.453
16.548
14.427
0.972
-91.869
18.058
78.672

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：The Scripps Research Institute Molecular Screening Center 靶标：N/A

External ID: FBW7_ACT_ALPHA_1536_1X%ACT PRUN

Protocol: Assay Overview:
FBW7 assay principle. In this assay, either mutant or wild type (w.t.) FBW7 interact with phosphorylated cyclin E peptide (cycE~P), which will bring donor and acceptor beads into close proximity. Laser excitation of the donor beads converts oxygen to an excited singlet state. Reaction of the singlet oxygen with the acceptor beads further activates a chemiluminescence/fluorescence reaction within the same bead resulting in emitted light at 520-620 nm. Small molecule activators that enhance the mutant FBW7 interaction with the cycE~P decrease the distance of the acceptor beads, thus leading to increased signal being emitted signal.
Protocol Summary:
There are six steps in this 1536 well assay format which are listed in order. First, 2.5uL/well of a 2X working solution containing RLFbw7 [12.5nM final], Cyclin E peptide [12.5nM final], and Ni beads [5ug/mL final], in assay buffer (25mM Tris-HCl pH 7.4 + 100mM NaCl, 0.1% Tween-20, 5mM ?-Mercaptoethanol and 0.05% BSA) was dispensed into columns 1-44. Then 2.5uL/well of a 2X working solution containing WTFbw7 [12.5nM final], Cyclin E peptide [12.5nM final], and Ni beads [5ug/mL final], in assay buffer was dispensed into columns 45-48. Using the pintool transfer device 134nL of compound or control was added to each well. This achieved a nominal screening concentration of 26.1uM and 2.6% DMSO concentration. This was followed by the addition of SA beads to all wells at 5ug/mL final concentration in assay buffer. The assay was then incubated for 20 hours in a temperature controlled 25C environment followed by Alphascreen detection using the PerkinElmer EnVision.

The percent activation for each compound was calculated as follows:

100 *( ( Test_Compound - Median_Low_Control ) / ( Median_High_Control - Median_Low_Control ) )
Where:
Test_Compound is defined as wells containing RLFbw7 (mutant), cyclin E peptide and Nickel acceptor beads in the presence of test compound
High_Control is defined as wells containing WTFbw7 (wild type), cyclin E peptide and Nickel acceptor beads
Low_Control is defined as the median of the wells containing DMSO, RLFbw7 (mutant), cyclin E peptide and Nickel acceptor beads
PubChem Activity Outcome and Score:

A mathematical algorithm was used to determine active compounds. Two values were calculated: (1) the average percent activation of all compounds tested for the screen, and (2) three times their standard deviation. The sum of these two values was used as a cutoff parameter, i.e. any compound that exhibited greater percent activation than the cutoff parameter (1.85% in the case here) was declared active.
The reported PubChem Activity Score has been normalized to 100% observed primary inhibition. Negative % inhibition values are reported as activity score zero.
The activity score range for active compounds is 100-1, for inactive 1-0.
List of Reagents:
Ni Beads- PerkinEmer Lifesciences Cat#6760619R
RLFbw7-Assay Provider
WTFbw7-Assay Provider
Cyclin E peptide-Assay Provider
5M NaCl- Sigma Cat# S6546-1L
Tween20- Fisher Cat# BP337
Tris 1M pH7.4 Research Organics Cat# 9686T
BSA-Sigma Cat#A7030
?-Mercaptoethanol-SigmaM6250
1536-well plates (Corning, part 7254)

Comment: Due to the size of the Scripps Molecular Screening Center compound library, this assay may have been run as two or more separate campaigns, each campaign testing a unique set of compounds. All data reported were normalized on a per-plate basis. Possible artifacts of this assay can include, but are not limited to: dust or lint located in or on wells of the microtiter plate, compounds that modulate well fluorescence. All test compound concentrations reported above and below are nominal; the specific test concentration(s) for a particular compound may vary based upon the actual sample provided by the Scripps Molecular Screening Center.

Inhibition at 26.1 uM
1.22
1.22
1.22
1.22
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21
1.21

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Trypanosoma brucei brucei

External ID: CHEMBL1109909

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: Bioorg. Med. Chem.
Year: 2010
Volume: 18
Issue: 7
First Page: 2566
Last Page: 2574
DOI: 10.1016/j.bmc.2010.02.034

Target ChEMBL ID: CHEMBL612851
ChEMBL Target Name: Trypanosoma brucei brucei
ChEMBL Target Type: ORGANISM - Target is a complete organism
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
0.03	IC100	=	0.03	uM
0.11	EC50	=	110	nM
1.3	EC50	=	1300	nM
5.7	IC50	=	5700	nM
50	IC50	>	50000	nM
8.5	IC50	=	8500	nM
35.5	IC50	=	35500	nM
50	IC50	>	50000	nM
50	IC50	>	50000	nM
50	IC50	>	50000	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Protein farnesyltransferase subunit beta

External ID: CHEMBL680085

Protocol: N/A

Comment: Journal: Bioorg. Med. Chem. Lett.
Year: 2003
Volume: 13
Issue: 21
First Page: 3661
Last Page: 3663
DOI: 10.1016/j.bmcl.2003.08.022

Target ChEMBL ID: CHEMBL2094108
ChEMBL Target Name: Protein farnesyltransferase
ChEMBL Target Type: PROTEIN COMPLEX - Target is a defined protein complex, consisting of multiple subunits
Relationship Type: D - Direct protein target assigned
Confidence: Direct protein complex subunits assigned

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units	Data Validity Comment
100	IC50	>	100000	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Histone-lysine N-methyltransferase SUV39H1

External ID: CHEMBL2173370

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: 2012
Volume: 56
First Page: 179
Last Page: 194
DOI: 10.1016/j.ejmech.2012.08.010

Target ChEMBL ID: CHEMBL1795118
ChEMBL Target Name: Histone-lysine N-methyltransferase SUV39H1
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
0.53	IC50	=	530	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Histone-lysine N-methyltransferase EHMT2

External ID: CHEMBL2173371

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: 2012
Volume: 56
First Page: 179
Last Page: 194
DOI: 10.1016/j.ejmech.2012.08.010

Target ChEMBL ID: CHEMBL6032
ChEMBL Target Name: Histone-lysine N-methyltransferase, H3 lysine-9 specific 3
ChEMBL Target Type: SINGLE PROTEIN - Target is a single protein chain
Relationship Type: H - Homologous protein target assigned
Confidence: Homologous single protein target assigned

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units
0.63	IC50	=	630	nM

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ICCB-Longwood/NSRB Screening Facility, Harvard Medical School 靶标：ORF 73 [Human herpesvirus 8 type M]

External ID: HMS791

Protocol: Prior to screening, FITC LANA1-23 was stored lyophilized at -80 degC and freshly purified chicken nucleosomes were stored at 4 degC in 20 mM Tris pH 7.5, 600 mM NaCl, 0.2 mM EDTA, 0.5 mM B-Mercaptoethanol.

On the day of screening, FITC LANA1-23 was resuspended at 50 uM in TEN-BT buffer (10 mM Tris-HCl (pH 7.5), 1 mM EDTA (pH 8.0), 2.5 mM NaCl, 5 mM Beta-mercaptoethanol, 0.01% Triton-X-100) and diluted to a final concentration of 50 nM in TEN-BT buffer plus 240 nM of purified nucleosomes (480 nM LANA peptide binding sites). 30 uL per well were dispensed in column 1-22 in Corning #3575 black 384 well plates.

Wells in column 23 contained 30 uL of the same mixture (for pilot screen) or with the addition of 10 uM monensin (for HTS) as a negative control. In column 24, 1250 nM unlabeled WT LANA1-23 peptide (for pilot screen) or 10 uM mitoxantrone (for HTS) was added to the wells as a positive control. Compounds were transferred into wells via stainless steel pin array (100 nL) and the reaction was incubated at room temperature for 10 to 45 minutes (stable for up to 2 hours). Library plates were screened in duplicate, with both assay plates in a given set prepared on the same day.

Following a room temperature incubation of 10 to 45 minutes, the assay is read on a EnVision plate reader using a 480 nM excitation filter, 535 nM S and P emission filters and D505fp/D535 dichoric mirror. mP value for FP measurement = 1000*(S-G*P)/(S+G*P) where S= , P=, G= G-factor. The G Factor = 1.

Comment: LANA 1-23 peptide containing the first 23 amino acids of the LANA protein from Kaposi's sarcoma herpesvirus (KSHV) was synthesized with an N-terminal FITC via a beta alanine linker and HPLC purified (peptide sequence: [FITC]-Beta alanine-MAPPGMRLRSGRSTGAPLTRGSC-[NH2]).

Data analysis: Z-scores were calculated for each replicate well using the mean and standard deviation of plate experimental well FP values. Compounds were considered active if the Z-score for both replicates < -2. Wells with high total fluorescence intensity (high S and P channel values) were excluded from further consideration. Activity scores were calculated based on replicate average FP Z-scores. For wells with average Z-score >= 0, the activity score was set to 0. For wells with replicate average Z-score < 0, replicate Z-score was divided by 4 and multiplied by -100. The replicate average was then used to determine the well activity score. Values > 100 (replicate average Z-score < -4) were set to 100.

FP_A	FP_B	Z-score_A	Z-score_B
91.8	90.2	0.126	0.03
93.2	94.9	0.261	0.457
94.8	92.9	0.416	0.275
89.9	85.7	-0.058	-0.379
91.5	90.1	0.097	0.021
88.7	89.6	-0.174	-0.025
92.5	88.2	0.194	-0.152
91.7	91.6	0.116	0.157
93.3	91.2	0.271	0.121
90.5	86.6	0	-0.297
87.1	88.8	-0.329	-0.097
87.6	85.8	-0.28	-0.37
89.2	87.4	-0.126	-0.224
87.2	86	-0.319	-0.351
85.8	86.1	-0.454	-0.342
88	92	-0.242	0.193
95.4	97.7	0.474	0.711
98.6	97.5	0.784	0.693
90	88.4	-0.048	-0.134
98.6	93.6	0.784	0.338

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：SH-SY5Y

External ID: CHEMBL3757989

Protocol: N/A

Comment: Journal: Eur. J. Med. Chem.
Year: 2016
Volume: 108
First Page: 553
Last Page: 563
DOI: 10.1016/j.ejmech.2015.12.003

Target ChEMBL ID: CHEMBL614910
ChEMBL Target Name: SH-SY5Y
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

Standard Type	Activity Comment
Activity	Active

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：N/A

External ID: CHEMBL4396795

Protocol: N/A

Comment: Journal: Bioorg Med Chem
Year: 2019
Volume: 27
Issue: 18
First Page: 4174
Last Page: 4184
DOI: 10.1016/j.bmc.2019.07.047

Standard Type	Standard Relation	Standard Value	Standard Units
Activity	>	100	%
Activity	=	6.1	%
Activity	>	100	%
Activity	=	11	%

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：Jurkat

External ID: CHEMBL1109912

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: Bioorg. Med. Chem.
Year: 2010
Volume: 18
Issue: 7
First Page: 2566
Last Page: 2574
DOI: 10.1016/j.bmc.2010.02.034

Target ChEMBL ID: CHEMBL397
ChEMBL Target Name: Jurkat
ChEMBL Target Type: CELL-LINE - Target is a specific cell-line
Relationship Type: N - Non-molecular target assigned
Confidence: Target assigned is non-molecular

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units	Activity Comment	Data Validity Comment
94.9	IC50	=	94900	nM
	IC50				Not Determined
141	IC50	=	141000	nM		Outside typical range
56	IC50	=	56000	nM
	IC50				Not Determined
	IC50				Not Determined
	IC50				Not Determined

HepG2 Cytotoxicity Assay Measured in Cell-Based System Using Plate Reader - 7071-02_Inhibitor_Dose_DryPowder_Activity_Set16

来源：ChEMBL 靶标：N/A

External ID: CHEMBL1109911

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: Bioorg. Med. Chem.
Year: 2010
Volume: 18
Issue: 7
First Page: 2566
Last Page: 2574
DOI: 10.1016/j.bmc.2010.02.034

PubChem Standard Value	Standard Type	Standard Relation	Standard Value	Standard Units	Activity Comment
0.27	IC50	=	270	nM
15	IC50	=	15000	nM
50	IC50	>	50000	nM
	IC50				Not Determined
50	IC50	>	50000	nM
50	IC50	>	50000	nM
	IC50				Not Determined
	IC50				Not Determined
	IC50				Not Determined

67-99-2 靶点实验数据