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BioAssay: AID 588477

Dose Response of round 1 SAR compounds for Cdc42 probe extension project tested by multiplex of eight GTPase proteins under the condition of Mg Buffer, non-chelator buffer

Assay Implementation: Lin Hong, Ph.D., J. Jacob Strouse, Ph.D., Mark Carter, M.S., Anna Waller, Ph.D. ..more
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 Tested Compounds
 Tested Compounds
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Active(1)
 
 
Inactive(4)
 
 
 Tested Substances
 Tested Substances
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Active(1)
 
 
Inactive(4)
 
 
AID: 588477
Data Source: NMMLSC (UNMCMD_Cd42PEC_MgSAR1_Dose_Response_Panel)
BioAssay Type: Panel
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-10-11
Hold-until Date: 2012-10-03
Modify Date: 2012-10-03

Data Table ( Complete ):           Active    All
BioActive Compound: 1
Depositor Specified Assays
AIDNameTypeComment
1772Project utilizing multiplex HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPasessummarySummary report for GTPase target project
761HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypescreeningHTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtype
Description:
University of New Mexico Assay Overview:
Assay Support: NIH I RO3 MH081231-01
HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases
PI: Angela Wandinger-Ness, Ph.D.
Co-PI: Larry Sklar, Ph.D.
Assay Development: Zurab Surviladze, Ph.D.
Assay Implementation: Lin Hong, Ph.D., J. Jacob Strouse, Ph.D., Mark Carter, M.S., Anna Waller, Ph.D.
Target Team Leader for the Center: Larry Sklar (lsklar@salud.unm.edu)
UNM Cheminformatics: Cristian Bologa, Ph.D., Oleg Ursu, Ph.D.

Chemistry: University of Kansas Specialized Chemistry Center
KU Specialized Chemistry Center PI: Jeff Aube, Ph.D.
KU SCC Project Manager: Jennifer E. Golden. Ph.D.
KU SCC Chemists on this project: Chad Schroeder, M.S., Denise Simpson, Ph.D., Julica Noeth, B.S.

Dose Response Assay Background and Significance:

Ras and related small molecular weight GTPases function in the regulation of signaling and cell growth, and collectively serve to control cell proliferation, differentiation and apoptosis [Tekai et al. 2001; Wennerberg et al. 2005]. The Ras-related GTPases are divided into four subfamilies with the Rab proteins regulating membrane transport, Rho proteins (including Rac and Cdc 42) regulating cytoskeletal rearrangements and responses to signaling, Arf/Sar proteins regulating membrane and microtubule dynamics as well as protein transport, and Ran proteins controlling nucleocytoplasmic transport. This project focuses on representative Ras, Rho, and Rab family members to validate the approach for the identification of new chemical compounds with novel therapeutic potential in cell signaling and growth control.

Ras and Ras-related GTPase functions are tightly regulated, and dysregulation is causal in a wide variety of human diseases. Ras mutations resulting in impaired GTP hydrolysis and plasma membrane hyperactivation are linked to many human cancers [Farnsworth et al. 1991; Sukumar et al. 1983; Taparowsky et al. 1982; Boylan et al. 1990; Hruban et al. 2004; Abrams et al. 1996]. Point mutations in the Rab and Rho GTPases are also causal in diverse human diseases affecting pigmentation, immune, and neurologic functions [Houlden et al. 2004; Verhoeven et al 2003; Williams et al. 2000; Bahaderan et al. 2003; and preliminary findings]. Rab and Rho mutants identified in human disease act as dominant negatives either due to a failure to bind GTP or due to inappropriate coupling of the active proteins with downstream effectors. To date, inhibition of Ras and Ras-related proteins has relied largely on altering membrane recruitment with various drugs affecting prenylation [Morgillo F and Lee HY, 2006; Russell RG, 2006; Park, et al. 2002]. Generally, Ras proteins must be farnesylated for proper membrane localization, while Rab and Rho proteins are geranylated. Such strategies lack specificity and are problematic because each of these prenylation machineries is required for the proper function of many Ras superfamily members. Rational drug design has only recently been applied to identify the first two small molecule inhibitors of Rho GTPase family members [Gao, et al. 2004; Nassar et al. 2006]. Therefore, broadly testing the Ras-related GTPases as targets for small molecule inhibitors and activators is expected to identify new classes of compounds that may be useful in the treatment of human disease, as well as in unraveling the molecular details of how Ras-related GTPases function.

The primary HTS assay was a no-wash fluorescent GTP-binding assay adapted to multiplexed, high-throughput measurements whereby multiple GTPases were simultaneously screened against the MLSCN library. The specificity is based on the observation that individual GTPases including wt and activated forms exhibit measurably distinct affinities for Bodipy-FI-GTP vs GTP. The assay involves the binding of fluorescent GTP to G protein-GST fusion proteins on GSH beads. A set of eight G proteins (Rab7 wt, Rac wt, Rac act, Ras wt, Ras act, Rho wt, Cdc42 wt, Cdc42 act) are arrayed under conditions of non-chelator buffer. Here we report the activity in Mg buffer conditions of various compounds that were synthesised by KU SCC under the Cdc42 Probe Extension Characterization project.
Panel Information
Panel dose response of eight GTPase proteins in Mg Buffer for Cdc42 probe extension project
    Data Table(Active)    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1Cdc42ACT32cell division cycle 42 (GTP binding protein, 25kDa) [Homo sapiens] [gi:56202836]
cell division cycle 42 activated mutant (GTP binding protein, 25kDa) [Homo sapiens]Taxonomy id: 9606
2Cdc42WT41cell division cycle 42 (GTP binding protein, 25kDa) [Homo sapiens] [gi:56202836]
cell division cycle 42 (GTP binding protein, 25kDa) [Homo sapiens]Taxonomy id: 9606
3Rab723GTP-binding protein (rab7) [Canis lupus familiaris] [gi:164058]
GTP-binding protein (rab7) [Canis lupus familiaris]Taxonomy id: 9615
4RacACT14Rac1 protein [Homo sapiens] [gi:8574038]
rac GTPase-activating protein 1 activated mutant [Homo sapiens]Taxonomy id: 9606
5RacWT14Rac1 protein [Homo sapiens] [gi:8574038]
rac GTPase-activating protein 1 [Homo sapiens]Taxonomy id: 9606
6RasACT5ras protein [Homo sapiens] [gi:190938]
ras protein activated mutant [Homo sapiens]Taxonomy id: 9606
7RasWT5ras protein [Homo sapiens] [gi:190938]
ras protein [Homo sapiens]Taxonomy id: 9606
8RhoWT14GTPase [Homo sapiens] [gi:407699]
transforming protein RhoA precursor [Homo sapiens]Taxonomy id: 9606

§ Panel component ID.
Protocol
Bead sets are coated with individual GST-small G proteins, blocked with Buffer (0.01% NP-40; 30mM HEPES pH 7.5; 100mM KCl; 20mM NaCl; 1mM MgCl2; 0.1% BSA and 1mM DTT), incubated overnight at 4 degrees C, and finally washed in buffer. The different bead sets, acquired from Duke Scientific, have similar size (~ 4 microm diameter) however they are distinguished by varied magnitude of emission at 665 +/-10 nm with excitation at 635 nm.

Test compounds were serially diluted 1:3 eight times for a total of nine different test compound concentrations in DMSO. Final compound dilutions in DMSO ranged from 1 microM to 10mM. These dilutions were then diluted 1 to 100 to give an assay concentration range of 10 nanoM to 100 microM.

The assay is conducted in 384-well microplates in a total well volume of 10.1 microliters (5 microliters of bead mixture, 0.1 microliters of test compound, and 5 microliters of 200 nanoM Bodipy-FL-GTP in buffer containing BSA and DTT for a final concentration of GTP of 100 nanoM). Negative Controls, which contain bead mixture and fluorescent GTP but no test compound, are located in columns 1 and 13 on plate. Positive Controls containing bead mixture with fluorescent GTP and 0.5 mM unlabeled GTP, are collected separately (either on teh same plate or different plate). Plates are placed on rotators and incubated for 40-45 minutes at 4 degrees C. Dose response experiments reported here include one multiplex format for: Rab7 wt, H-Ras wt, H-Ras constitutively active, Rac wt, Rac constitutively active, Rho wt, Cdc42 wt, and Cdc42 constitutively active.

Sample acquisition and preliminary analysis is conducted with the HyperCyt(R) high throughput flow cytometry platform. The HyperCyt system interfaces a flow cytometer and autosampler for high-throughput microliter volume sampling from 384-well microtiter plates [Kuckuck, et al., 2001; Ramirez, et al., 2003]. The stream of particles is excited at 488 nm and 635 nm, and flow cytometric data of light scatter and fluorescence emission at 530 +/- 20 nm (FL1) and emission at 665 +/- 10 nm (FL8) are collected on a Cyan Flow Cytometer (Dako). Analysis of the time-resolved acquisition data file uses HyperView software to merge the flow cytometry data files with compound worklist files generated by HyperSip software. Gating based on forward scatter (FS) and side scatter (SS) parameters is used to identify singlet bead populations. Gating based on FL8 emission distinguishes the beads coated with different proteins, and the green median fluorescence intensity (MFI) per bead population (well) is calculated. The raw data are parsed in HyperView to produce annotated fluorescence summary data for each well. The parsed data are then processed through an Excel template file constructed specifically for the assay to fit the data via GraphPad Prism.

Calculations:

In dose response experiments, the percent response was calculated for each GTPase target by the following equation:
%Response = (SampleMFI - PCntrl)/(NCntrl - PCntrl)
where SampleMFI is the median fluorescence intensity of the compound sample, PCntrl is the median fluorescence intensity measured in the presence of access blocking non-fluorescent GTP, and NCntrl is the median fluorescence intensity measured in the presence of DMSO. The %Response was 100% for compounds not affecting the binding of Bodipy-FL-GTP, and greater than 100% if the compound increased the binding of Bodipy-FL-GTP and less than 100% if the compound decreased the binding of Bodipy-FL-GTP.

The % response values for the entire concentration range of a test compound were fitted by Prism(R) software (GraphPad Software, Inc., San Diego, CA) using nonlinear least-squares regression in a sigmoidal dose response model with variable slope, also known as the four parameter logistic equation. Curve fit statistics were used to determine the following parameters of the model: EC50, microM - concentration of added test compound competitor that inhibited fluorescent ligand binding by 50 percent; LOGEC50 - the logarithm of EC50; TOP - the response value at the top plateau; BOTTOM - the response value at the bottom plateau; HILLSLOPE - the slope factor, or the Hill coefficient; STD_LOGEC50, STD_TOP, STD_BOTTOM, STD_HILLSLOPE - standard errors of LOGEC50, TOP, BOTTOM, and HILLSLOPE ; EC50_95CI_LOW, EC50_95CI_HIGH - the low and high boundaries of the 95% confidence interval of the EC50 estimate, RSQR - the correlation coefficient (r squared) indicative of goodness-of-fit.

Assessment of the validity of the estimates from the Prism fits was filtered via the following criteria:
- -8 < LOGEC50 < -4 (the computed EC50 value should be in the interval of tested concentrations)
- STD_LOGEC50/|LOGEC50| <0.15 (the standard error of LOGEC50 should be no greater than 15% of the absolute value of LOGEC50)
- 0.5 < |HILLSLOPE| < 2 (the absolute value of HILLSLOPE should be higher than 0.5 and lower than 2)

PUBCHEM_ACTIVITY_SCORE were calculated based on an EC50 cutoff of 10 microM, by using the following equations:
SCORE = 100*(1-EC50/10 )
And compounds were demeaned active if the EC50 is equal to or less than 10 microM and the span of the %Response is greater than or equal to 30%. Span is the magnitude of %Response change over the tested concentration range based on either fitted values of TOP minus BOTTOM or from %Response values at lowest to highest concentration.

An active compound have a PUBCHEM_ACTIVITY_SCORE greater than 0.

Keywords: NIH Roadmap, NMMLSC, high throughput flow cytometry, Rab7 wt, Ras wt, Ras activated, Rac wt, Rac activated, Rho wt, Cdc42 wt, Cdc42 activated, multiplex, bead-based, screening, dose response, SAR, UNMCMD
Categorized Comment
BAO: version: 1.4b1080

BAO: bioassay specification: assay stage: lead optimization

BAO: bioassay specification: assay biosafety level: bsl1

BAO: bioassay specification: assay measurement type: endpoint assay

BAO: bioassay specification: assay readout content: assay readout method: regular screening

BAO: bioassay specification: assay readout content: content readout type: multiplexed readout

BAO: meta target: molecular target: protein target: enzyme: protease

BAO: meta target detail: binding reporter specification: interaction: protein-nucleotide

BAO: detection technology: fluorescence: flow cytometry

Result Definitions
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TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Cdc42ACT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target1cell division cycle 42 (GTP binding protein, 25kDa) [Homo sapiens]Outcome
2Cdc42ACT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target1Integer
3Cdc42ACT_ACTIVITY_TYPEQualifier for the value of EC501String
4Cdc42ACT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit1FloatμM
5Cdc42ACT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate1FloatμM
6Cdc42ACT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate1FloatμM
7Cdc42ACT_BOTTOMResponse value at the bottom plateau1Float%
8Cdc42ACT_TOPResponse value at the top plateau1Float%
9Cdc42ACT_LOGEC50Log of the EC50 estimate1Float
10Cdc42ACT_HILLSLOPEHill slope estimate for the fitted dose response curve1Float
11Cdc42ACT_STD_BOTTOMStandard error for the response value at the bottom plateau1Float%
12Cdc42ACT_STD_TOPStandard error for the response value at the top plateau1Float%
13Cdc42ACT_STD_LOGEC50Standard error for the Log EC50 value1Float
14Cdc42ACT_STD_HILLSLOPEStandard error for the HILL SLOPE1Float
15Cdc42ACT_RSQRCorrelation coefficient for the fitted dose response curve1Float
16Cdc42ACT_N_POINTSNumber of data points for each dose response curve1Integer
17Cdc42ACT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition1Float%
18Cdc42ACT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition1Float%
19Cdc42ACT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition1Float%
20Cdc42ACT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition1Float%
21Cdc42ACT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition1Float%
22Cdc42ACT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition1Float%
23Cdc42ACT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition1Float%
24Cdc42ACT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition1Float%
25Cdc42ACT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition1Float%
26Cdc42ACT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition1Float%
27Cdc42ACT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition1Float%
28Cdc42ACT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition1Float%
29Cdc42ACT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition1Float%
30Cdc42ACT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition1Float%
31Cdc42ACT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition1Float%
32Cdc42ACT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition1Float%
33Cdc42ACT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition1Float%
34Cdc42ACT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition1Float%
35Cdc42ACT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition1Float%
36Cdc42ACT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition1Float%
37Cdc42WT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target2cell division cycle 42 (GTP binding protein, 25kDa) [Homo sapiens]Outcome
38Cdc42WT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target2Integer
39Cdc42WT_ACTIVITY_TYPEQualifier for the value of EC502String
40Cdc42WT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit2FloatμM
41Cdc42WT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate2FloatμM
42Cdc42WT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate2FloatμM
43Cdc42WT_BOTTOMResponse value at the bottom plateau2Float%
44Cdc42WT_TOPResponse value at the top plateau2Float%
45Cdc42WT_LOGEC50Log of the EC50 estimate2Float
46Cdc42WT_HILLSLOPEHill slope estimate for the fitted dose response curve2Float
47Cdc42WT_STD_BOTTOMStandard error for the response value at the bottom plateau2Float%
48Cdc42WT_STD_TOPStandard error for the response value at the top plateau2Float%
49Cdc42WT_STD_LOGEC50Standard error for the Log EC50 value2Float
50Cdc42WT_STD_HILLSLOPEStandard error for the HILL SLOPE2Float
51Cdc42WT_RSQRCorrelation coefficient for the fitted dose response curve2Float
52Cdc42WT_N_POINTSNumber of data points for each dose response curve2Integer
53Cdc42WT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition2Float%
54Cdc42WT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition2Float%
55Cdc42WT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition2Float%
56Cdc42WT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition2Float%
57Cdc42WT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition2Float%
58Cdc42WT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition2Float%
59Cdc42WT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition2Float%
60Cdc42WT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition2Float%
61Cdc42WT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition2Float%
62Cdc42WT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition2Float%
63Cdc42WT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition2Float%
64Cdc42WT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition2Float%
65Cdc42WT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition2Float%
66Cdc42WT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition2Float%
67Cdc42WT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition2Float%
68Cdc42WT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition2Float%
69Cdc42WT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition2Float%
70Cdc42WT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition2Float%
71Cdc42WT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition2Float%
72Cdc42WT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition2Float%
73Rab7_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target3GTP-binding protein (rab7) [Canis lupus familiaris]Outcome
74Rab7_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target3Integer
75Rab7_ACTIVITY_TYPEQualifier for the value of EC503String
76Rab7_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit3FloatμM
77Rab7_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate3FloatμM
78Rab7_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate3FloatμM
79Rab7_BOTTOMResponse value at the bottom plateau3Float%
80Rab7_TOPResponse value at the top plateau3Float%
81Rab7_LOGEC50Log of the EC50 estimate3Float
82Rab7_HILLSLOPEHill slope estimate for the fitted dose response curve3Float
83Rab7_STD_BOTTOMStandard error for the response value at the bottom plateau3Float%
84Rab7_STD_TOPStandard error for the response value at the top plateau3Float%
85Rab7_STD_LOGEC50Standard error for the Log EC50 value3Float
86Rab7_STD_HILLSLOPEStandard error for the HILL SLOPE3Float
87Rab7_RSQRCorrelation coefficient for the fitted dose response curve3Float
88Rab7_N_POINTSNumber of data points for each dose response curve3Integer
89Rab7_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition3Float%
90Rab7_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition3Float%
91Rab7_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition3Float%
92Rab7_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition3Float%
93Rab7_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition3Float%
94Rab7_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition3Float%
95Rab7_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition3Float%
96Rab7_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition3Float%
97Rab7_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition3Float%
98Rab7_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition3Float%
99Rab7_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition3Float%
100Rab7_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition3Float%
101Rab7_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition3Float%
102Rab7_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition3Float%
103Rab7_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition3Float%
104Rab7_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition3Float%
105Rab7_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition3Float%
106Rab7_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition3Float%
107Rab7_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition3Float%
108Rab7_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition3Float%
109RacACT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target4Rac1 protein [Homo sapiens]Outcome
110RacACT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target4Integer
111RacACT_ACTIVITY_TYPEQualifier for the value of EC504String
112RacACT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit4FloatμM
113RacACT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate4FloatμM
114RacACT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate4FloatμM
115RacACT_BOTTOMResponse value at the bottom plateau4Float%
116RacACT_TOPResponse value at the top plateau4Float%
117RacACT_LOGEC50Log of the EC50 estimate4Float
118RacACT_HILLSLOPEHill slope estimate for the fitted dose response curve4Float
119RacACT_STD_BOTTOMStandard error for the response value at the bottom plateau4Float%
120RacACT_STD_TOPStandard error for the response value at the top plateau4Float%
121RacACT_STD_LOGEC50Standard error for the Log EC50 value4Float
122RacACT_STD_HILLSLOPEStandard error for the HILL SLOPE4Float
123RacACT_RSQRCorrelation coefficient for the fitted dose response curve4Float
124RacACT_N_POINTSNumber of data points for each dose response curve4Integer
125RacACT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition4Float%
126RacACT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition4Float%
127RacACT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition4Float%
128RacACT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition4Float%
129RacACT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition4Float%
130RacACT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition4Float%
131RacACT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition4Float%
132RacACT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition4Float%
133RacACT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition4Float%
134RacACT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition4Float%
135RacACT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition4Float%
136RacACT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition4Float%
137RacACT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition4Float%
138RacACT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition4Float%
139RacACT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition4Float%
140RacACT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition4Float%
141RacACT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition4Float%
142RacACT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition4Float%
143RacACT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition4Float%
144RacACT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition4Float%
145RacWT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target5Rac1 protein [Homo sapiens]Outcome
146RacWT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target5Integer
147RacWT_ACTIVITY_TYPEQualifier for the value of EC505String
148RacWT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit5FloatμM
149RacWT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate5FloatμM
150RacWT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate5FloatμM
151RacWT_BOTTOMResponse value at the bottom plateau5Float%
152RacWT_TOPResponse value at the top plateau5Float%
153RacWT_LOGEC50Log of the EC50 estimate5Float
154RacWT_HILLSLOPEHill slope estimate for the fitted dose response curve5Float
155RacWT_STD_BOTTOMStandard error for the response value at the bottom plateau5Float%
156RacWT_STD_TOPStandard error for the response value at the top plateau5Float%
157RacWT_STD_LOGEC50Standard error for the Log EC50 value5Float
158RacWT_STD_HILLSLOPEStandard error for the HILL SLOPE5Float
159RacWT_RSQRCorrelation coefficient for the fitted dose response curve5Float
160RacWT_N_POINTSNumber of data points for each dose response curve5Integer
161RacWT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition5Float%
162RacWT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition5Float%
163RacWT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition5Float%
164RacWT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition5Float%
165RacWT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition5Float%
166RacWT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition5Float%
167RacWT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition5Float%
168RacWT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition5Float%
169RacWT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition5Float%
170RacWT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition5Float%
171RacWT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition5Float%
172RacWT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition5Float%
173RacWT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition5Float%
174RacWT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition5Float%
175RacWT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition5Float%
176RacWT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition5Float%
177RacWT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition5Float%
178RacWT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition5Float%
179RacWT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition5Float%
180RacWT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition5Float%
181RasACT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target6ras protein [Homo sapiens]Outcome
182RasACT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target6Integer
183RasACT_ACTIVITY_TYPEQualifier for the value of EC506String
184RasACT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit6FloatμM
185RasACT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate6FloatμM
186RasACT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate6FloatμM
187RasACT_BOTTOMResponse value at the bottom plateau6Float%
188RasACT_TOPResponse value at the top plateau6Float%
189RasACT_LOGEC50Log of the EC50 estimate6Float
190RasACT_HILLSLOPEHill slope estimate for the fitted dose response curve6Float
191RasACT_STD_BOTTOMStandard error for the response value at the bottom plateau6Float%
192RasACT_STD_TOPStandard error for the response value at the top plateau6Float%
193RasACT_STD_LOGEC50Standard error for the Log EC50 value6Float
194RasACT_STD_HILLSLOPEStandard error for the HILL SLOPE6Float
195RasACT_RSQRCorrelation coefficient for the fitted dose response curve6Float
196RasACT_N_POINTSNumber of data points for each dose response curve6Integer
197RasACT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition6Float%
198RasACT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition6Float%
199RasACT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition6Float%
200RasACT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition6Float%
201RasACT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition6Float%
202RasACT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition6Float%
203RasACT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition6Float%
204RasACT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition6Float%
205RasACT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition6Float%
206RasACT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition6Float%
207RasACT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition6Float%
208RasACT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition6Float%
209RasACT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition6Float%
210RasACT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition6Float%
211RasACT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition6Float%
212RasACT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition6Float%
213RasACT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition6Float%
214RasACT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition6Float%
215RasACT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition6Float%
216RasACT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition6Float%
217RasWT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target7ras protein [Homo sapiens]Outcome
218RasWT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target7Integer
219RasWT_ACTIVITY_TYPEQualifier for the value of EC507String
220RasWT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit7FloatμM
221RasWT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate7FloatμM
222RasWT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate7FloatμM
223RasWT_BOTTOMResponse value at the bottom plateau7Float%
224RasWT_TOPResponse value at the top plateau7Float%
225RasWT_LOGEC50Log of the EC50 estimate7Float
226RasWT_HILLSLOPEHill slope estimate for the fitted dose response curve7Float
227RasWT_STD_BOTTOMStandard error for the response value at the bottom plateau7Float%
228RasWT_STD_TOPStandard error for the response value at the top plateau7Float%
229RasWT_STD_LOGEC50Standard error for the Log EC50 value7Float
230RasWT_STD_HILLSLOPEStandard error for the HILL SLOPE7Float
231RasWT_RSQRCorrelation coefficient for the fitted dose response curve7Float
232RasWT_N_POINTSNumber of data points for each dose response curve7Integer
233RasWT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition7Float%
234RasWT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition7Float%
235RasWT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition7Float%
236RasWT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition7Float%
237RasWT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition7Float%
238RasWT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition7Float%
239RasWT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition7Float%
240RasWT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition7Float%
241RasWT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition7Float%
242RasWT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition7Float%
243RasWT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition7Float%
244RasWT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition7Float%
245RasWT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition7Float%
246RasWT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition7Float%
247RasWT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition7Float%
248RasWT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition7Float%
249RasWT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition7Float%
250RasWT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition7Float%
251RasWT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition7Float%
252RasWT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition7Float%
253RhoWT_PUBCHEM_ACTIVITY_OUTCOMEOutcome of compound for this protein target8GTPase [Homo sapiens]Outcome
254RhoWT_PUBCHEM_ACTIVITY_SCOREScore rating for this protein target8Integer
255RhoWT_ACTIVITY_TYPEQualifier for the value of EC508String
256RhoWT_EC50_MICROMEffective concentration of half maximal event count as estimated by curve fit8FloatμM
257RhoWT_EC50_95CI_LOWLower 95% confidence interval boundary for the EC50 curve fit estimate8FloatμM
258RhoWT_EC50_95CI_HIGHUpper 95% confidence interval boundary for the EC50 curve fit estimate8FloatμM
259RhoWT_BOTTOMResponse value at the bottom plateau8Float%
260RhoWT_TOPResponse value at the top plateau8Float%
261RhoWT_LOGEC50Log of the EC50 estimate8Float
262RhoWT_HILLSLOPEHill slope estimate for the fitted dose response curve8Float
263RhoWT_STD_BOTTOMStandard error for the response value at the bottom plateau8Float%
264RhoWT_STD_TOPStandard error for the response value at the top plateau8Float%
265RhoWT_STD_LOGEC50Standard error for the Log EC50 value8Float
266RhoWT_STD_HILLSLOPEStandard error for the HILL SLOPE8Float
267RhoWT_RSQRCorrelation coefficient for the fitted dose response curve8Float
268RhoWT_N_POINTSNumber of data points for each dose response curve8Integer
269RhoWT_%Response_0uM_Rep1 (0μM**)Percent Response measured with 0 micromolar concentration of test compound first repetition8Float%
270RhoWT_%Response_0.015uM_Rep1 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound first repetition8Float%
271RhoWT_%Response_0.04uM_Rep1 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound first repetition8Float%
272RhoWT_%Response_0.14uM_Rep1 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound first repetition8Float%
273RhoWT_%Response_0.41uM_Rep1 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound first repetition8Float%
274RhoWT_%Response_1.23uM_Rep1 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound first repetition8Float%
275RhoWT_%Response_3.7uM_Rep1 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound first repetition8Float%
276RhoWT_%Response_11.2uM_Rep1 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound first repetition8Float%
277RhoWT_%Response_33.1uM_Rep1 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound first repetition8Float%
278RhoWT_%Response_100uM_Rep1 (100μM**)Percent Response measured with 100 micromolar concentration of test compound first repetition8Float%
279RhoWT_%Response_0uM_Rep2 (0μM**)Percent Response measured with 0 micromolar concentration of test compound second repetition8Float%
280RhoWT_%Response_0.015uM_Rep2 (0.015μM**)Percent Response measured with 0.015 micromolar concentration of test compound second repetition8Float%
281RhoWT_%Response_0.04uM_Rep2 (0.04μM**)Percent Response measured with 0.04 micromolar concentration of test compound second repetition8Float%
282RhoWT_%Response_0.14uM_Rep2 (0.14μM**)Percent Response measured with 0.14 micromolar concentration of test compound second repetition8Float%
283RhoWT_%Response_0.41uM_Rep2 (0.41μM**)Percent Response measured with 0.41 micromolar concentration of test compound second repetition8Float%
284RhoWT_%Response_1.23uM_Rep2 (1.23μM**)Percent Response measured with 1.23 micromolar concentration of test compound second repetition8Float%
285RhoWT_%Response_3.7uM_Rep2 (3.7μM**)Percent Response measured with 3.7 micromolar concentration of test compound second repetition8Float%
286RhoWT_%Response_11.2uM_Rep2 (11.2μM**)Percent Response measured with 11.2 micromolar concentration of test compound second repetition8Float%
287RhoWT_%Response_33.1uM_Rep2 (33.1μM**)Percent Response measured with 33.1 micromolar concentration of test compound second repetition8Float%
288RhoWT_%Response_100uM_Rep2 (100μM**)Percent Response measured with 100 micromolar concentration of test compound second repetition8Float%

** Test Concentration. § Panel component ID.
Additional Information
Grant Number: NIH I RO3 MH081231-01

Classification
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