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BioAssay: AID 624129

Dose Response confirmation of uHTS hits from a small molecule antagonists of the CXCR6 receptor in a screening assay

Prostate cancer (PCa) is the second leading cause of cancer death in American men and its morbidity has increased globally in recent years. The high mortality rate is closely associated with the spread of malignant cells to various tissues including bone. Nearly 10% of patients whose conditions are diagnosed as PCa initially present with bone metastasis and almost all patients who die of prostate more ..
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 Tested Compounds
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Active(15)
 
 
Inactive(595)
 
 
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Active(15)
 
 
Inactive(595)
 
 
AID: 624129
Data Source: Burnham Center for Chemical Genomics (SBCCG-A845-CXCR6-DMSO-Panel-Assay)
BioAssay Type: Panel, Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-05-08

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 15
Related Experiments
Show more
AIDNameTypeComment
602244uHTS identification of CXCR6 Inhibitors in a B-arrestin luminescence assayScreeningdepositor-specified cross reference
602249Summary assay for small molecule Inhibitors of CXCR6Summarydepositor-specified cross reference
602409Single concentration confirmation of uHTS hits from a small molecule antagonists of the CXCR6 receptor via a luminescent beta-arrestin assayScreeningsame project related to Summary assay
651594Dose Response counterscreen of small molecule antagonists of the CXCR6 receptor using a CXCR5 receptor luminescent beta-arrestin assayConfirmatorysame project related to Summary assay
651598SAR analysis of small molecule antagonists of the CXCR6 receptor in an APJ-DiscoveRx counter screen panel assayConfirmatorysame project related to Summary assay
651601SAR analysis of small molecule antagonists of the CXCR6 receptor in a screening panel assayConfirmatorysame project related to Summary assay
651708SAR analysis of small molecule antagonists of the CXCR6 receptor in a screening panel assay - 3Confirmatorysame project related to Summary assay
651902SAR analysis of small molecule antagonists of the CXCR6 receptor using a CXCR4 receptor luminescent beta-arrestin counterscreenConfirmatorysame project related to Summary assay
651919SAR analysis of small molecule antagonists of the CXCR6 receptor in a screening panel assay - 4Confirmatorysame project related to Summary assay
651940SAR analysis of small molecule antagonists of the hCXCR6 receptor using a mCXCR6 receptor luminescent beta-arrestin selectivity assayConfirmatorysame project related to Summary assay
651941SAR analysis of small molecule antagonists of the CXCR6 receptor using a CCR6 receptor luminescent beta-arrestin counterscreenConfirmatorysame project related to Summary assay
Description:
Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG)
Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA)
Network: NIH Molecular Libraries Probe Production Centers Network (MLPCN)
Grant Number: 1R03MH095589-01 (Cycle 18)
Assay Provider: Gregory Roth Ph.D., Sanford Burnham Medical Research Institute.

Prostate cancer (PCa) is the second leading cause of cancer death in American men and its morbidity has increased globally in recent years. The high mortality rate is closely associated with the spread of malignant cells to various tissues including bone. Nearly 10% of patients whose conditions are diagnosed as PCa initially present with bone metastasis and almost all patients who die of prostate cancers have skeletal involvement. Identifying new mechanisms that control bone metastasis is of great consequence to facilitate the design of therapeutics aimed at decreasing metastatic risk and/or its complications. To address this unmet medical need, our team is actively engaged in exploring the chemical biology, medicinal chemistry, and therapeutic significance of modulating tumor cell trafficking and metastasis via chemokine receptor inhibition. The primary objective of this proposal is to use high throughput screening methods to identify small molecule antagonist probes that selectively inhibit CXCR6. Our team intends to address a key hypothesis: The CXCR6/CXCL16 axis significantly contributes to PCa cell metastasis and subsequent bone invasion. A small molecule antagonist would block cancer cell trafficking; hence mediate a metastatic event and disease progression to bone. Thus, access to pharmacologically available small molecule antagonists will ultimately enable our studies in disease relevant models and allow for a more seamless translational advance to clinical applications.

The project goal is to identify a chemical probe of chemokine CXCR6 receptor function that inhibits the receptor. An antagonist of this receptor would provide a novel research tool to evaluate cancer cell trafficking and bone metastasis in prostrate and other cancers

In this description we utilize enzyme-fragment complementation to directly measure GPCR activation. Unlike imaging or other second messenger assays, the DiscoveRx b-Arrestin assay allows for a direct measure of GPCR activation by detection of b-Arrestin binding to the CXCR6 receptor. In this system, b-Arrestin is fused to an N-terminal deletion mutant of b-gal (termed the enzyme acceptor of EA) and the GPCR of interest is fused to a smaller (42 amino acids), weakly complementing fragment termed ProLink. In cells that stably express these fusion proteins, ligand stimulation results in the interaction of b-Arrestin and the Prolink-tagged GPCR, forcing the complementation of the two b-gal fragments and resulting in the formation of a functional enzyme that converts substrate to detectable signal.

This assay is a follow-up to "uHTS identification of CXCR6 Inhibitors in a B-arrestin luminescence assay", AID 602244. Compounds were tested in the primary assay against the CXCR6 receptor. Compounds considered active in this assay were further tested in the same B-arrestin system but against the APJ and CCR6 receptors. Both receptor assays were employed to test selectivity of compounds for the CXCR6 chemokine receptor over the CCR6 chemokine receptor and an unrelated GPCR. In addition, compounds were tested in a b-galactosidase enzyme assay to test for compounds being potential artifacts in the primary assays based on b-galactosidase detection. Compounds considered active in the CCR6, APJ and B-galactosidase assays are non-selective, non-specific and/or assay artifacts of the primary assay and are not desired.

REFERENCES
Hu, W; Zhen, X; Xiong, B; Wang, B; Zhang, W; Zhou, W CXCR6 is expressed in human prostate cancer in vivo and is involved in the in vitro invasion of PC3 and LNCap cells. Cancer Sci 2008, 99, 1362-1369.

Matloubian, M; David, A; Engel, S; Ryan, JE; Cyster, JG A transmembrane CXC chemokine is a ligand for HIV-coreceptor Bonzo. Nature Immunol 2000, 1, 298-304.

Chandrasekar B, Bysani S, Mummidi S. CXCL16 signals via Gi, phosphatidylinositol 3-kinase, Akt, I kappa B kinase, and nuclear factor-kappa B and induces cell-cell adhesion and aortic smooth muscle cell proliferation. J Biol Chem 2004, 279, 3188-3196.
Panel Information
panel file
    Data Table(Active)    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescription
ActiveInactive
1IC50_1_Range1_APJ-Discoverx36417apelin receptor [Homo sapiens] [gi:4885057]
IC50 value determined using a sigmoidal dose response equation
2IC50_2_Range1_APJ-Discoverxapelin receptor [Homo sapiens] [gi:4885057]
IC50 value determined using a sigmoidal dose response equation
3IC50_3_Range1_APJ-Discoverxapelin receptor [Homo sapiens] [gi:4885057]
IC50 value determined using a sigmoidal dose response equation
4IC50_1_Range1_b-galactosidase44337RecName: Full=Beta-galactosidase; Short=Beta-gal; AltName: Full=Lactase [gi:75404427]
IC50 value determined using a sigmoidal dose response equation
5IC50_1_Range1_CCR63747C-C chemokine receptor type 6 [Homo sapiens] [gi:37187860]
IC50 value determined using a sigmoidal dose response equation
6IC50_2_Range1_CCR6C-C chemokine receptor type 6 [Homo sapiens] [gi:37187860]
IC50 value determined using a sigmoidal dose response equation
7IC50_1_Range1_CXCR6241369C-X-C chemokine receptor type 6 [Homo sapiens] [gi:5730106]
IC50 value determined using a sigmoidal dose response equation
8IC50_1_Range2_CXCR6C-X-C chemokine receptor type 6 [Homo sapiens] [gi:5730106]
IC50 value determined using a sigmoidal dose response equation
9IC50_2_Range1_CXCR6C-X-C chemokine receptor type 6 [Homo sapiens] [gi:5730106]
IC50 value determined using a sigmoidal dose response equation
10IC50_2_Range2_CXCR6C-X-C chemokine receptor type 6 [Homo sapiens] [gi:5730106]
IC50 value determined using a sigmoidal dose response equation

§ Panel component ID.
Protocol
1)
A. Brief Description of the Assay:
The purpose of this assay is to detect antagonists that inhibit the activation of the Angiotensin II receptor-like 1 (Apelin receptor) in the CHO-K1 AGTRL-1 beta-Arrestin Cell Line in 1536-well plate format in uHTS mode.
B. Materials:
Angiotensin II receptor-like 1 (AGTRL-1) Cell Line (DiscoveRx, Cat# 93-0250C2)
F12 nutrient mix HAMs (Invitrogen, Cat# 11765)
Fetal Bovine Serum, heat-inactivated (Hyclone, Cat# SH30396)
100X Penicillin/Streptomycin Solution (Invitrogen, Cat#15140-122)
Hygromycin B (Roche, Cat# 10843555001)
Geneticin (MPBiomedicals, Cat # 1672548)
Trypsin-EDTA 0.25% (Invitrogen, Cat# 25200-056)
Cell Dissociation Buffer (Invitrogen, Cat# 13151)
DPBS (Hyclone, Cat# 30028.02)
T225 TC Flask (Nunc, Cat# 159934)
Cell strainer, 40 um (BD, Cat# 352340)
1536-well, white, solid-bottom, Kalypsys compatible, TC plate (Corning)
Apelin-13 (Sigma-Aldrich, Cat# A6469)
PathHunter Detection Reagents (DiscoveRx, Cat# 93-0001)
Galacton Star
Emerald 11
Cell Assay Buffer
C. uHTS Procedures:
Day1 Cell Seeding
1) Plate 1000 cells/well in 4 uL of assay media into columns 1-48 of a 1536-well assay plate, using straight tip dispense on a Kalypsys dispenser.
2) Centrifuge plates at 500 rpm for 1 minute on a Vspin centrifuge. Use Kalypsys metal lids.
3) Incubate overnight at 37 degrees, 100% relative humidity, 5% CO2 for 16-18 hours.
Day2 Compound Addition
1) Centrifuge compound plates at 500 rpm for 1 minute on a Vspin centrifuge.
2) Using LabCyte Echo, transfer 60 nL from a 2 mM Echo qualified plate containing test compounds into assay plate Col. 5 - 48 (final concentration of test compounds is 20 uM, 1% DMSO). Transfer 60 nL of DMSO to positive and negative control wells in Columns 1 - 4.
3) Immediately following compound/DMSO transfer via the Echo, using the Kalypsys Dispenser, transfer 2ul/well of Assay media to Col. 1-2 for the positive control wells.
4) Using the Kalypsys Dispenser, add 2ul/well of 30 nM Apelin-13 (FAC = 10 nM) in assay media to Col. 3-48 for the negative control and test compound wells .
5) Centrifuge plates at 500 rpm for 1 minute on a Vspin centrifuge.
6) Incubate plates at 25 degrees in the dark for 90 minutes.
7) Following 90 minute incubation, deliver 3.0 uL of Detection Reagent solution to each assay plate (Columns 1 - 48) using a Kalypsys dispenser.
8) Centrifuge plates at 2000 rpm for 3 minute on a Vspin centrifuge.
9) Incubate plates for 60 minutes at 25 degrees in the dark.
10) Read plates using the Viewlux using a luminescence protocol.
D. Recipes:
Growth Media
F12 nutrient mix HAMs supplemented with 10% hi-FBS, 1X Penicillin/Streptomycin; selection reagents: 300ug/ml Hygromycin B, 800ug/ml Geneticin
Assay Media
Same as Growth Media without the selection reagents
Trypsin
Dilute 0.25% Trypsin/EDTA to 0.05% Trypsin/EDTA using DPBS
Positive Control
Growth Media with 30 nM Apelin-13
Detection Reagent
Use the following ratio to prepare the detection reagent:
Galacton Star : Emerald II : Assay Buffer = 1 : 5 : 19
2)
A. Brief Description of the Assay:
The purpose of this assay is to detect activators of beta-galactosidase in 1536-well plate format in uHTS mode.
B: Assay materials:
1) Beta-Galactosidase from Sigma-Aldrich (Cat# G4155)
2) Assay Medium: Opti-MEM Medium supplemented with 1% hiFBS, 1X Pen/Strep/Glu, 125 ug/mL Hygromycin (1/2 recommended), 250 ug/mL Geneticin (1/2 recommended)
3) DiscoverX b-Arrestin Detection Reagents: Galacton Star, Emerald II, and Cell Assay Buffer
Dose Response protocol:
1) Add 5.0 uL of b-gal diluted in assay media to 0.03 U/mL to all wells (0.02 U/mL final assay concentration) of a 1536-well plate with the exception of the positive control wells where 5.0 uL of assay media alone is added.
2) Spin plates at 500 rpm for 1 min.
3) Prepare Detection Reagent Solution from DiscoveRx (1 part Galacton Star: 5 parts Emerald II and 19 parts Cell Assay Buffer).
4) Using a Labcyte Echo, DMSO and test compounds are transferred to assay wells. DMSO only is transferred to positive and negative control wells, while varying volumes of test compounds are transferred to test compound wells to achieve the desired test concentrations. Test compound wells in the assay plate are back-filled with DMSO to equalize final assay concentrations.
5) Spin plates at 500 rpm for 1 min.
6) Add 2.5 uL of Detection Reagent Solution to all wells of assay plate and immediately spin plates at 500 rpm for 1 min.
7) After 15 minutes, read assay plates on a Perkin Elmer Envision using a luminescence protocol
3)
A. Brief Description of the Assay:
The purpose of this assay is to detect antagonists that inhibit the activation of the CCR6 receptor in the CHO-K1 beta-Arrestin Cell Line in 1536-well plate format in uHTS mode.
B. Materials:
PathHunter CHO-K1 CCR6 b-arrestin cell line (DiscoveRx, Cat# 93-0194C2)
F12 nutrient mix HAMs (Invitrogen, Cat# 11765)
Fetal Bovine Serum, heat-inactivated (Hyclone, Cat# SH30396)
100X Penicillin/Streptomycin Solution (Invitrogen, Cat#15140-122)
Hygromycin B (Roche, Cat# 10843555001)
Geneticin (MPBiomedicals, Cat # 1672548)
Trypsin-EDTA 0.25% (Invitrogen, Cat# 25200-056)
Cell Dissociation Buffer (Invitrogen, Cat# 13151)
DPBS (Hyclone, Cat# 30028.02)
T225 TC Flask (Nunc, Cat# 159934)
1536-well, white, solid-bottom, Kalypsys compatible, TC plate (Aurora)
MIP-3a/CCL20 peptide (R&D Systems, Cat# 360MP)
PathHunter Detection Reagents (DiscoveRx, Cat# 93-0001)
Galacton Star
Emerald 11
Cell Assay Buffer
C. uHTS Procedures:
Day1 Cell Seeding
1) Plate 800 cells/well in 4 uL of assay media into columns 1-48 of a 1536-well assay plate, using Biotek dispenser.
2) Centrifuge plates at 500 rpm for 1 minute on a Vspin centrifuge. Use Kalypsys metal lids.
3) Incubate overnight at 37 degrees, 100% relative humidity, 5% CO2 for 16-18 hours.
Day2 Compound Addition
1) Centrifuge compound plates at 500 rpm for 1 minute on a Vspin centrifuge.
2) Using LabCyte Echo 555, transfer 40 nL of DMSO to positive and negative control wells in columns 1 - 4 and 45-48, respectively. Using a dose response protocol, transfer compounds from 10mM and 0.312 mM Echo qualified plates into assay plate columns 5 - 45. (Final concentrations range 66 uM to 0.128 uM, 10 doses, with 0.66% DMSO.)
3) Immediately following compound/DMSO transfer via the Echo, using the Biotek Dispenser, transfer 2ul/well of Assay media to Col. 1-2 for the positive control wells.
4) Using the Biotek Dispenser, add 2ul/well of 42 nM MIP3a/CCL20 (FAC = 14 nM) in assay media to Col. 3-48 for the negative control and test compound wells.
5) Centrifuge plates at 500 rpm for 1 minute on a Vspin centrifuge.
6) Incubate plates at 25 degrees in the dark for 90 minutes.
7) Following 90 minute incubation, deliver 3.0 uL of Detection Reagent solution to each assay plate (Columns 1 - 48) using a Biotek dispenser.
8) Centrifuge plates at 2000 rpm for 2 minute on a Vspin centrifuge.
9) Incubate plates for 60 minutes at 25 degrees in the dark.
10) Read plates using the Envision using a luminescence protocol.
D. Recipes:
Growth Media
F12 nutrient mix HAMs supplemented with 10% hi-FBS, 1X Penicillin/Streptomycin; selection reagents: 300ug/ml Hygromycin B, 800ug/ml Geneticin
Assay Media/Positive Control
Same as Growth Media without the selection reagents and only 2.5% hi-FBS
Trypsin
Dilute 0.25% Trypsin/EDTA to 0.05% Trypsin/EDTA using DPBS
Negative Control
Growth Media with 14 nM MIP3a/CCL20
Detection Reagent
Use the following ratio to prepare the detection reagent:
Galacton Star : Emerald II : Assay Buffer = 1 : 5 : 19
4)
A. Brief Description of the CXCR6 Assay:
The purpose of this assay is to detect antagonists that cause the inhibit the Chemokine CXCR6 receptor in the CHO-K1 PathHunter CHO-K1 CXCR6 b-arrestin cell line in 1536-well plate format.
B. Materials:
PathHunter CHO-K1 CXCR6 b-arrestin cell line (DiscoveRx, Cat# 93-0205C2)
F12 nutrient mix HAMs (Invitrogen, Cat# 11765)
Fetal Bovine Serum, heat-inactivated (Hyclone, Cat# SH30396)
100X Penicillin/Streptomycin Solution (Invitrogen, Cat#15140-122)
Hygromycin B (Roche, Cat# 10843555001)
Geneticin (MPBiomedicals, Cat# 1672548)
Trypsin-EDTA 0.25% (Invitrogen, Cat# 25200-056)
Cell Dissociation Buffer (Invitrogen, Cat# 13151)
DPBS (Hyclone, Cat# 30028.02)
T225 TC Flask (Nunc, Cat# 159934)
Cell strainer, 40 um (BD, Cat# 352340)
1536-well, white, solid-bottom, Kalypsys compatible, TC plate (Corning)
CXCCL16 (R&D Systems, Cat# 976-CX)
PathHunter Detection Reagents (DiscoveRx, Cat# 93-0001)
Galacton Star
Emerald 11
Cell Assay Buffer
C. uHTS Procedures:
Day1 Cell Seeding
1) Plate 800 cells/well in 3 uL of assay media into columns 1-48 of a 1536-well assay plate, using combi dispenser.
2) Centrifuge plates at 500 rpm for 1 minute on a Vspin centrifuge.
3) Put Kalypsis lids on and incubate overnight at 37 degrees, 100% relative humidity, 5% CO2 for 16-18 hours.
Day2 Compound Addition
1) Centrifuge compound plates at 500 rpm for 1 minute on a Vspin centrifuge.
2) Using LabCyte Echo, transfer 40-2.5 nl from a 10 mM and 0.312 mM Echo qualified plate containing DPI liquid reordered test into assay plate Col. 5 - 44 (final concentration of test compounds is 79uM to 0.156 uM, 0.8% DMSO). Add 40nl DMSOt to positive and negative control wells in Columns 1 - 4 and 45-48.
3) Immediately following compound/DMSO transfer via the Echo, using the Biotek Dispenser, transfer 2ul/well of Assay media to Col. 1-2 for the positive control.
4) Using the Biotek Dispenser, add 2ul/well of 25 nM CXCL16 (FAC = 10 nM) in assay media to Col. 3-48 for the negative control and test compounds.
5) Centrifuge plates at 1000 rpm for 1 minute on a Vspin centrifuge.
6) Incubate plates at 25 degrees in the dark for 90 minutes.
7) Following 90 minute incubation, deliver 3.0 uL of Detection Reagent solution to each assay plate (Columns 1 - 48) using a Biotek dispenser.
8) Centrifuge plates at 2000 rpm for 3 minute on a Vspin centrifuge.
9) Incubate plates for 60 minutes at 25 degrees in the dark.
10) Read plates using the Envision plate reader using a luminescence protocol.
D. Recipes:
Growth Media
F12 nutrient mix HAMs supplemented with 10% hi-FBS, 1X Penicillin/Streptomycin; selection reagents: 300ug/ml Hygromycin B, 800ug/ml Geneticin
Assay Media
Same as Growth Media without the selection reagents
Trypsin
Dilute 0.25% Trypsin/EDTA to 0.05% Trypsin/EDTA using DPBS
Negative Control
Growth Media with 10 nM CXCL16
Detection Reagent
Use the following ratio to prepare the detection reagent:
Galacton Star : Emerald II : Assay Buffer = 1 : 5 : 19
Comment
Compounds that were active in the CXCR6 assay, IC50 < 50 uM were tested in 3 counterscreens. Compounds were considered active in the counter screens as follows:
B-galactosidase IC50 < 50uM

APJ IC50 < 5 * IC50 CXCR6 result

CCR6 IC50 < 5 * IC50 CXCR6 result
Compounds that were found to be active in the CXCR6 assay and not active in the counter screens are considered "active" in this panel. The score is the value from the CXCR6 assay if the compound is active, otherwise the panel score is 41.
To simplify the distinction between the inactives of the primary screen and of the confirmatory screening stage, the Tiered Activity Scoring System was developed and implemented.
Activity Scoring
Activity scoring rules were devised to take into consideration compound efficacy, its potential interference with the assay and the screening stage that the data was obtained. Details of the Scoring System will be published elsewhere. Briefly, the outline of the scoring system utilized for the assay is as follows:
1) First tier (0-40 range) is reserved for primary screening data and is not applicable in this assay
2) Second tier (41-80 range) is reserved for dose-response confirmation data
a. Inactive compounds of the confirmatory stage are assigned a score value equal 41.
b. The score is linearly correlated with a compound's potency and, in addition, provides a measure of the likelihood that the compound is not an artifact based on the available information.
c. The Hill coefficient is taken as a measure of compound behavior in the assay via an additional scaling factor QC:
QC = 2.6*[exp(-0.5*nH^2) - exp(-1.5*nH^2)]
This empirical factor prorates the likelihood of target- or pathway-specific compound effect vs. its non-specific behavior in the assay. This factor is based on expectation that a compound with a single mode of action that achieved equilibrium in the assay demonstrates the Hill coefficient value of 1. Compounds deviating from that behavior are penalized proportionally to the degree of their deviation.
d. Summary equation that takes into account all the items discussed above is
Score = 44 + 6*(pIC50-3)*QC,
Where pIC50 is a negative log(10) of the IC50 value expressed in mole/L concentration units. This equation results in the Score values above 50 for compounds that demonstrate high potency and predictable behavior. Compounds that are inactive in the assay or whose concentration-dependent behavior are likely to be an artifact of that assay will generally have lower Score values.
3) Third tier (81-100 range) is reserved for resynthesized true positives and their analogues and is not applicable in this assay
Result Definitions
Show more
TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PUBCHEM_ACTIVITY_SCORE_APJ-DiscoverxThe Bioassay Ranking Score1apelin receptor [Homo sapiens]Integer
2Outcome_APJ-DiscoverxOutcome of the assay1Outcome
3IC50_APJ-Discoverx_Mean_QualifierThis qualifier is to be used with the next TID, IC50_Mean. If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value1String
4IC50_APJ-Discoverx_MeanIC50 value determined using a sigmoidal dose response equation1FloatμM
5PUBCHEM_ACTIVITY_SCORE_b-galactosidaseThe Bioassay Ranking Score4RecName: Full=Beta-galactosidase; Short=Beta-gal; AltName: Full=LactaseInteger
6Outcome_b-galactosidaseOutcome of the assay4Outcome
7IC50_B-galactosidase_Mean_QualifierThis qualifier is to be used with the next TID, IC50_Mean. If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value4String
8IC50_B-galactosidase_MeanIC50 value determined using a sigmoidal dose response equation4FloatμM
9PUBCHEM_ACTIVITY_SCORE_CCR6The Bioassay Ranking Score5C-C chemokine receptor type 6 [Homo sapiens]Integer
10Outcome_CCR6Outcome of the assay5Outcome
11IC50_CCR6_Mean_QualifierThis qualifier is to be used with the next TID, IC50_Mean. If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value7C-X-C chemokine receptor type 6 [Homo sapiens]String
12IC50_CCR6_MeanIC50 value determined using a sigmoidal dose response equation7FloatμM
13PUBCHEM_ACTIVITY_SCORE_CXCR6The Bioassay Ranking Score7Integer
14Outcome_CXCR6Outcome of the assay7Outcome
15IC50_CXCR6_Mean_QualifierThis qualifier is to be used with the next TID, IC50_Mean. If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that valueString
16IC50_CXCR6_Mean*IC50 value determined using a sigmoidal dose response equationFloatμM
17IC50_Qualifier_1_Range1_APJ-DiscoverxThis qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value7C-X-C chemokine receptor type 6 [Homo sapiens]String
18IC50_1_Range1_APJ-DiscoverxIC50 value determined using a sigmoidal dose response equation1apelin receptor [Homo sapiens]FloatμM
19Std.Err(IC50)_1_Range1_APJ-DiscoverxStandard Error of the IC50 valueFloatμM
20nH_1_Range1_APJ-DiscoverxHill coefficient determined using sigmoidal dose response equationFloat
21Excluded_Points_first_point_Range1_APJ-DiscoverxFlags to indicate which of the first dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
22% Activity at 80 uM_first_point_Range1_APJ-Discoverx (80μM**)% Activity at the test concentrationFloat%
23% Activity at 40 uM_first_point_Range1_APJ-Discoverx (40μM**)% Activity at the test concentrationFloat%
24% Activity at 20 uM_first_point_Range1_APJ-Discoverx (20μM**)% Activity at the test concentrationFloat%
25% Activity at 10 uM_first_point_Range1_APJ-Discoverx (10μM**)% Activity at the test concentrationFloat%
26% Activity at 5 uM_first_point_Range1_APJ-Discoverx (5μM**)% Activity at the test concentrationFloat%
27% Activity at 2.5 uM_first_point_Range1_APJ-Discoverx (2.5μM**)% Activity at the test concentrationFloat%
28% Activity at 1.25 uM_first_point_Range1_APJ-Discoverx (1.25μM**)% Activity at the test concentrationFloat%
29% Activity at 0.625 uM_first_point_Range1_APJ-Discoverx (0.625μM**)% Activity at the test concentrationFloat%
30% Activity at 0.3125 uM_first_point_Range1_APJ-Discoverx (0.3125μM**)% Activity at the test concentrationFloat%
31% Activity at 0.15625 uM_first_point_Range1_APJ-Discoverx (0.15625μM**)% Activity at the test concentrationFloat%
32IC50_Qualifier_2_Range1_APJ-DiscoverxThis qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value2apelin receptor [Homo sapiens]String
33IC50_2_Range1_APJ-DiscoverxIC50 value determined using a sigmoidal dose response equation2FloatμM
34Std.Err(IC50)_2_Range1_APJ-DiscoverxStandard Error of the IC50 valueFloatμM
35nH_2_Range1_APJ-DiscoverxHill coefficient determined using sigmoidal dose response equationFloat
36Excluded_Points_second_point_Range1_APJ-DiscoverxFlags to indicate which of the second dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
37% Activity at 80 uM_second_point_Range1_APJ-Discoverx (80μM**)% Activity at the test concentrationFloat%
38% Activity at 40 uM_second_point_Range1_APJ-Discoverx (40μM**)% Activity at the test concentrationFloat%
39% Activity at 20 uM_second_point_Range1_APJ-Discoverx (20μM**)% Activity at the test concentrationFloat%
40% Activity at 10 uM_second_point_Range1_APJ-Discoverx (10μM**)% Activity at the test concentrationFloat%
41% Activity at 5 uM_second_point_Range1_APJ-Discoverx (5μM**)% Activity at the test concentrationFloat%
42% Activity at 2.5 uM_second_point_Range1_APJ-Discoverx (2.5μM**)% Activity at the test concentrationFloat%
43% Activity at 1.25 uM_second_point_Range1_APJ-Discoverx (1.25μM**)% Activity at the test concentrationFloat%
44% Activity at 0.625 uM_second_point_Range1_APJ-Discoverx (0.625μM**)% Activity at the test concentrationFloat%
45% Activity at 0.3125 uM_second_point_Range1_APJ-Discoverx (0.3125μM**)% Activity at the test concentrationFloat%
46% Activity at 0.15625 uM_second_point_Range1_APJ-Discoverx (0.15625μM**)% Activity at the test concentrationFloat%
47IC50_Qualifier_3_Range1_APJ-DiscoverxThis qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value3apelin receptor [Homo sapiens]String
48IC50_3_Range1_APJ-DiscoverxIC50 value determined using a sigmoidal dose response equation3FloatμM
49Std.Err(IC50)_3_Range1_APJ-DiscoverxStandard Error of the IC50 valueFloatμM
50nH_3_Range1_APJ-DiscoverxHill coefficient determined using sigmoidal dose response equationFloat
51Excluded_Points_third_point_Range1_APJ-DiscoverxFlags to indicate which of the third dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
52% Activity at 80 uM_third_point_Range1_APJ-Discoverx (80μM**)% Activity at the test concentrationFloat%
53% Activity at 40 uM_third_point_Range1_APJ-Discoverx (40μM**)% Activity at the test concentrationFloat%
54% Activity at 20 uM_third_point_Range1_APJ-Discoverx (20μM**)% Activity at the test concentrationFloat%
55% Activity at 10 uM_third_point_Range1_APJ-Discoverx (10μM**)% Activity at the test concentrationFloat%
56% Activity at 5 uM_third_point_Range1_APJ-Discoverx (5μM**)% Activity at the test concentrationFloat%
57% Activity at 2.5 uM_third_point_Range1_APJ-Discoverx (2.5μM**)% Activity at the test concentrationFloat%
58% Activity at 1.25 uM_third_point_Range1_APJ-Discoverx (1.25μM**)% Activity at the test concentrationFloat%
59% Activity at 0.625 uM_third_point_Range1_APJ-Discoverx (0.625μM**)% Activity at the test concentrationFloat%
60% Activity at 0.3125 uM_third_point_Range1_APJ-Discoverx (0.3125μM**)% Activity at the test concentrationFloat%
61% Activity at 0.15625 uM_third_point_Range1_APJ-Discoverx (0.15625μM**)% Activity at the test concentrationFloat%
62IC50_Qualifier_1_Range1_b-galactosidaseThis qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value4RecName: Full=Beta-galactosidase; Short=Beta-gal; AltName: Full=LactaseString
63IC50_1_Range1_b-galactosidaseIC50 value determined using a sigmoidal dose response equation4FloatμM
64Std.Err(IC50)_1_Range1_b-galactosidaseStandard Error of the IC50 valueFloatμM
65nH_1_Range1_b-galactosidaseHill coefficient determined using sigmoidal dose response equationFloat
66Excluded_Points_first_point_Range1_b-galactosidaseFlags to indicate which of the first dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
67% Activity at 80 uM_first_point_Range1_b-galactosidase (80μM**)% Activity at the test concentrationFloat%
68% Activity at 40 uM_first_point_Range1_b-galactosidase (40μM**)% Activity at the test concentrationFloat%
69% Activity at 20 uM_first_point_Range1_b-galactosidase (20μM**)% Activity at the test concentrationFloat%
70% Activity at 10 uM_first_point_Range1_b-galactosidase (10μM**)% Activity at the test concentrationFloat%
71% Activity at 5 uM_first_point_Range1_b-galactosidase (5μM**)% Activity at the test concentrationFloat%
72% Activity at 2.5 uM_first_point_Range1_b-galactosidase (2.5μM**)% Activity at the test concentrationFloat%
73% Activity at 1.25 uM_first_point_Range1_b-galactosidase (1.25μM**)% Activity at the test concentrationFloat%
74% Activity at 0.625 uM_first_point_Range1_b-galactosidase (0.625μM**)% Activity at the test concentrationFloat%
75% Activity at 0.3125 uM_first_point_Range1_b-galactosidase (0.3125μM**)% Activity at the test concentrationFloat%
76% Activity at 0.15625 uM_first_point_Range1_b-galactosidase (0.15625μM**)% Activity at the test concentrationFloat%
77IC50_Qualifier_1_Range1_CCR6This qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value5C-C chemokine receptor type 6 [Homo sapiens]String
78IC50_1_Range1_CCR6IC50 value determined using a sigmoidal dose response equation5FloatμM
79Std.Err(IC50)_1_Range1_CCR6Standard Error of the IC50 valueFloatμM
80nH_1_Range1_CCR6Hill coefficient determined using sigmoidal dose response equationFloat
81Excluded_Points_first_point_Range1_CCR6Flags to indicate which of the first dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
82% Activity at 80 uM_first_point_Range1_CCR6 (80μM**)% Activity at the test concentrationFloat%
83% Activity at 40 uM_first_point_Range1_CCR6 (40μM**)% Activity at the test concentrationFloat%
84% Activity at 20 uM_first_point_Range1_CCR6 (20μM**)% Activity at the test concentrationFloat%
85% Activity at 10 uM_first_point_Range1_CCR6 (10μM**)% Activity at the test concentrationFloat%
86% Activity at 5 uM_first_point_Range1_CCR6 (5μM**)% Activity at the test concentrationFloat%
87% Activity at 2.5 uM_first_point_Range1_CCR6 (2.5μM**)% Activity at the test concentrationFloat%
88% Activity at 1.25 uM_first_point_Range1_CCR6 (1.25μM**)% Activity at the test concentrationFloat%
89% Activity at 0.625 uM_first_point_Range1_CCR6 (0.625μM**)% Activity at the test concentrationFloat%
90% Activity at 0.3125 uM_first_point_Range1_CCR6 (0.3125μM**)% Activity at the test concentrationFloat%
91% Activity at 0.15625 uM_first_point_Range1_CCR6 (0.15625μM**)% Activity at the test concentrationFloat%
92IC50_Qualifier_2_Range1_CCR6This qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value6C-C chemokine receptor type 6 [Homo sapiens]String
93IC50_2_Range1_CCR6IC50 value determined using a sigmoidal dose response equation6FloatμM
94Std.Err(IC50)_2_Range1_CCR6Standard Error of the IC50 valueFloatμM
95nH_2_Range1_CCR6Hill coefficient determined using sigmoidal dose response equationFloat
96Excluded_Points_second_point_Range1_CCR6Flags to indicate which of the second dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
97% Activity at 80 uM_second_point_Range1_CCR6 (80μM**)% Activity at the test concentrationFloat%
98% Activity at 40 uM_second_point_Range1_CCR6 (40μM**)% Activity at the test concentrationFloat%
99% Activity at 20 uM_second_point_Range1_CCR6 (20μM**)% Activity at the test concentrationFloat%
100% Activity at 10 uM_second_point_Range1_CCR6 (10μM**)% Activity at the test concentrationFloat%
101% Activity at 5 uM_second_point_Range1_CCR6 (5μM**)% Activity at the test concentrationFloat%
102% Activity at 2.5 uM_second_point_Range1_CCR6 (2.5μM**)% Activity at the test concentrationFloat%
103% Activity at 1.25 uM_second_point_Range1_CCR6 (1.25μM**)% Activity at the test concentrationFloat%
104% Activity at 0.625 uM_second_point_Range1_CCR6 (0.625μM**)% Activity at the test concentrationFloat%
105% Activity at 0.3125 uM_second_point_Range1_CCR6 (0.3125μM**)% Activity at the test concentrationFloat%
106% Activity at 0.15625 uM_second_point_Range1_CCR6 (0.15625μM**)% Activity at the test concentrationFloat%
107IC50_Qualifier_1_Range1_CXCR6This qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value7C-X-C chemokine receptor type 6 [Homo sapiens]String
108IC50_1_Range1_CXCR6IC50 value determined using a sigmoidal dose response equation7FloatμM
109Std.Err(IC50)_1_Range1_CXCR6Standard Error of the IC50 valueFloatμM
110nH_1_Range1_CXCR6Hill coefficient determined using sigmoidal dose response equationFloat
111Excluded_Points_first_point_Range1_CXCR6Flags to indicate which of the first dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
112% Activity at 80 uM_first_point_Range1_CXCR6 (80μM**)% Activity at the test concentrationFloat%
113% Activity at 40 uM_first_point_Range1_CXCR6 (40μM**)% Activity at the test concentrationFloat%
114% Activity at 20 uM_first_point_Range1_CXCR6 (20μM**)% Activity at the test concentrationFloat%
115% Activity at 10 uM_first_point_Range1_CXCR6 (10μM**)% Activity at the test concentrationFloat%
116% Activity at 5 uM_first_point_Range1_CXCR6 (5μM**)% Activity at the test concentrationFloat%
117% Activity at 2.5 uM_first_point_Range1_CXCR6 (2.5μM**)% Activity at the test concentrationFloat%
118% Activity at 1.25 uM_first_point_Range1_CXCR6 (1.25μM**)% Activity at the test concentrationFloat%
119% Activity at 0.625 uM_first_point_Range1_CXCR6 (0.625μM**)% Activity at the test concentrationFloat%
120% Activity at 0.3125 uM_first_point_Range1_CXCR6 (0.3125μM**)% Activity at the test concentrationFloat%
121% Activity at 0.15625 uM_first_point_Range1_CXCR6 (0.15625μM**)% Activity at the test concentrationFloat%
122IC50_Qualifier_1_Range2_CXCR6This qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value8C-X-C chemokine receptor type 6 [Homo sapiens]String
123IC50_1_Range2_CXCR6IC50 value determined using a sigmoidal dose response equation8FloatμM
124Std.Err(IC50)_1_Range2_CXCR6Standard Error of the IC50 valueFloatμM
125nH_1_Range2_CXCR6Hill coefficient determined using sigmoidal dose response equationFloat
126Excluded_Points_second_point_Range1_CXCR6Flags to indicate which of the second dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
127% Activity at 80 uM_second_point_Range1_CXCR6 (80μM**)% Activity at the test concentrationFloat%
128% Activity at 40 uM_second_point_Range1_CXCR6 (40μM**)% Activity at the test concentrationFloat%
129% Activity at 20 uM_second_point_Range1_CXCR6 (20μM**)% Activity at the test concentrationFloat%
130% Activity at 10 uM_second_point_Range1_CXCR6 (10μM**)% Activity at the test concentrationFloat%
131% Activity at 5 uM_second_point_Range1_CXCR6 (5μM**)% Activity at the test concentrationFloat%
132% Activity at 2.5 uM_second_point_Range1_CXCR6 (2.5μM**)% Activity at the test concentrationFloat%
133% Activity at 1.25 uM_second_point_Range1_CXCR6 (1.25μM**)% Activity at the test concentrationFloat%
134% Activity at 0.625 uM_second_point_Range1_CXCR6 (0.625μM**)% Activity at the test concentrationFloat%
135% Activity at 0.3125 uM_second_point_Range1_CXCR6 (0.3125μM**)% Activity at the test concentrationFloat%
136% Activity at 0.15625 uM_second_point_Range1_CXCR6 (0.15625μM**)% Activity at the test concentrationFloat%
137IC50_Qualifier_2_Range1_CXCR6This qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value9C-X-C chemokine receptor type 6 [Homo sapiens]String
138IC50_2_Range1_CXCR6IC50 value determined using a sigmoidal dose response equation9FloatμM
139Std.Err(IC50)_2_Range1_CXCR6Standard Error of the IC50 valueFloatμM
140nH_2_Range1_CXCR6Hill coefficient determined using sigmoidal dose response equationFloat
141Excluded_Points_first_point_Range2_CXCR6Flags to indicate which of the first dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
142% Activity at 79 uM_first_point_Range2_CXCR6 (79μM**)% Activity at the test concentrationFloat%
143% Activity at 39.5 uM_first_point_Range2_CXCR6 (39.5μM**)% Activity at the test concentrationFloat%
144% Activity at 19.75 uM_first_point_Range2_CXCR6 (19.75μM**)% Activity at the test concentrationFloat%
145% Activity at 9.875 uM_first_point_Range2_CXCR6 (9.875μM**)% Activity at the test concentrationFloat%
146% Activity at 4.9375 uM_first_point_Range2_CXCR6 (4.9375μM**)% Activity at the test concentrationFloat%
147% Activity at 2.46875 uM_first_point_Range2_CXCR6 (2.46875μM**)% Activity at the test concentrationFloat%
148% Activity at 1.234375 uM_first_point_Range2_CXCR6 (1.23438μM**)% Activity at the test concentrationFloat%
149% Activity at 0.6171875 uM_first_point_Range2_CXCR6 (0.617188μM**)% Activity at the test concentrationFloat%
150% Activity at 0.3085938 uM_first_point_Range2_CXCR6 (0.308594μM**)% Activity at the test concentrationFloat%
151% Activity at 0.1542969 uM_first_point_Range2_CXCR6 (0.154297μM**)% Activity at the test concentrationFloat%
152IC50_Qualifier_2_Range2_CXCR6This qualifier is to be used with the next TID, . If the qualifier is "=", the IC50 result equals the value in that column. If the qualifier is ">", the IC50 result is greater than that value. If the qualifier is "<", the IC50 result is smaller than that value10C-X-C chemokine receptor type 6 [Homo sapiens]String
153IC50_2_Range2_CXCR6IC50 value determined using a sigmoidal dose response equation10FloatμM
154Std.Err(IC50)_2_Range2_CXCR6Standard Error of the IC50 valueFloatμM
155nH_2_Range2_CXCR6Hill coefficient determined using sigmoidal dose response equationFloat
156Excluded_Points_second_point_Range2_CXCR6Flags to indicate which of the second dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
157% Activity at 79 uM_second_point_Range2_CXCR6 (79μM**)% Activity at the test concentrationFloat%
158% Activity at 39.5 uM_second_point_Range2_CXCR6 (39.5μM**)% Activity at the test concentrationFloat%
159% Activity at 19.75 uM_second_point_Range2_CXCR6 (19.75μM**)% Activity at the test concentrationFloat%
160% Activity at 9.875 uM_second_point_Range2_CXCR6 (9.875μM**)% Activity at the test concentrationFloat%
161% Activity at 4.9375 uM_second_point_Range2_CXCR6 (4.9375μM**)% Activity at the test concentrationFloat%
162% Activity at 2.46875 uM_second_point_Range2_CXCR6 (2.46875μM**)% Activity at the test concentrationFloat%
163% Activity at 1.234375 uM_second_point_Range2_CXCR6 (1.23438μM**)% Activity at the test concentrationFloat%
164% Activity at 0.6171875 uM_second_point_Range2_CXCR6 (0.617188μM**)% Activity at the test concentrationFloat%
165% Activity at 0.3085938 uM_second_point_Range2_CXCR6 (0.308594μM**)% Activity at the test concentrationFloat%
166% Activity at 0.1542969 uM_second_point_Range2_CXCR6 (0.154297μM**)% Activity at the test concentrationFloat%

* Activity Concentration. ** Test Concentration. § Panel component ID.
Additional Information
Grant Number: 1R03MH095589-01 (Cycle 18)

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