SAR analysis of small molecule antagonists of the APJ receptor via a luminescent beta-arrestin assay - BioAssay Summary
Currently there are no small molecule tools to investigate the biological functions of apelin and its receptor. Apelin is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) APJ (angiotensin II receptor-like 1, AGTRL-1 and APLNR). Until the discovery of apelin, APJ was an orphan GPCR. APJ is coupled to Gai, and has been shown in cell culture to inhibit adenylate cyclase. The more ..
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Active(25)
 
 
Inactive(31)
 
 
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Active(25)
 
 
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 Related BioAssays
 Related BioAssays
Table of Contents
AID: 463109
Data Source: Burnham Center for Chemical Genomics (BCCG-A404-APJ-Antagonist-DryPowder-Assay)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-08-19
Hold-until Date: 2011-08-18
Modify Date: 2011-08-18

Data Table (Complete):           Active    All
Target
BioActive Compounds: 25
Depositor Specified Assays
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AIDNameTypeComment
2521uHTS identification of small molecule antagonists of the APJ receptor via a luminescent beta-arrestin assayscreeningPrimary Screen
2569Summary assay for small molecule antagonists of the APJ receptorsummarySummary AID.
Description:
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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: 1R21NS059422-01
Assay Provider: Dr. Layton Smith, Sanford-Burnham Medical Research Institute

Currently there are no small molecule tools to investigate the biological functions of apelin and its receptor. Apelin is the endogenous peptide ligand for the G-protein coupled receptor (GPCR) APJ (angiotensin II receptor-like 1, AGTRL-1 and APLNR). Until the discovery of apelin, APJ was an orphan GPCR. APJ is coupled to Gai, and has been shown in cell culture to inhibit adenylate cyclase. The APJ gene encodes a receptor that most closely resembles the angiotensin receptor AT1. However, the APJ receptor does not bind angiotensin II. Underscoring the emerging importance of the apelin/APJ system, recent studies have shown that apelin reduces the extent of atherosclerotic lesions in ApoE-/- mice, and opposes the development of abdominal aortic aneurysms. Additional research has revealed that APJ forms a heterodimer with the Ang II receptor AT1, and that this complex facilitates antagonism of Ang II signaling by apelin. Despite these exciting results, there remains a multitude of unanswered questions regarding the role of apelin and APJ in physiology and pathology.
The project goal is to identify a chemical probe of apelin receptor function that transiently and reversibly activates the receptor. An antagonist or inhibitor receptor activation would provide a novel research tool to evaluate the role of apelin in cardiovascular and metabolic disease pathology.
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 APJ 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. Antagonists would be expected to inhibit agonist activation of the receptor resulting in the inhibition of signal formation in this assay.

This assay has been developed and performed to confirm hits originally identified in "uHTS identification of small molecule antagonists of the APJ receptor via a luminescent beta-arrestin assay" (AID 2521) and to study the structure-activity relationship on analogs of the confirmed hits. Compounds are either acquired from commercial sources or synthesized internally.
Protocol
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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

Dose Response 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 varying volumes from a 2 mM Echo qualified plate containing test compounds into assay plate Col. 5 - 48 to achieve the desired dose response concentrations and range. Backfill with DMSO to equalize final DMSO concentrations in all test wells. Transfer equal volume 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.

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
Comment
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Compounds with IC50_Mean < 10 uM are defined as "active" in this assay.

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. Its utilization for the assay is described below.

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 and is not applicable in this assay.

3) Third tier (81-100 range) is reserved for resynthesized true positives and their analogues and is not applicable in this assay
a. Inactive compounds of the confirmatory stage are assigned a score value equal 81.
b. The score is linearly correlated with a compound 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-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 LYP1 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 the items discussed above is
Score = 82 + 3*(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 85 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.
Result Definitions
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Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1IC50_Mean_QualifierThis qualifier is to be used with the next TID, IC50_Mean. If 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
2IC50_Mean*IC50 value determined using sigmoidal dose response equationFloatμM
3Excluded_Points_first_point_Range1Flags 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
4% Activity at 79 uM_first_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
5% Activity at 39.5 uM_first_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
6% Activity at 19.75 uM_first_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
7% Activity at 9.875 uM_first_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
8% Activity at 4.9375 uM_first_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
9% Activity at 2.46875 uM_first_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
10% Activity at 1.234375 uM_first_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
11% Activity at 0.6171875 uM_first_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
12% Activity at 0.3085938 uM_first_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
13% Activity at 0.1542969 uM_first_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
14Excluded_Points_second_point_Range1Flags 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
15% Activity at 79 uM_second_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
16% Activity at 39.5 uM_second_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
17% Activity at 19.75 uM_second_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
18% Activity at 9.875 uM_second_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
19% Activity at 4.9375 uM_second_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
20% Activity at 2.46875 uM_second_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
21% Activity at 1.234375 uM_second_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
22% Activity at 0.6171875 uM_second_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
23% Activity at 0.3085938 uM_second_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
24% Activity at 0.1542969 uM_second_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
25Excluded_Points_third_point_Range1Flags 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
26% Activity at 79 uM_third_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
27% Activity at 39.5 uM_third_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
28% Activity at 19.75 uM_third_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
29% Activity at 9.875 uM_third_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
30% Activity at 4.9375 uM_third_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
31% Activity at 2.46875 uM_third_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
32% Activity at 1.234375 uM_third_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
33% Activity at 0.6171875 uM_third_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
34% Activity at 0.3085938 uM_third_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
35% Activity at 0.1542969 uM_third_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
36Excluded_Points_fourth_point_Range1Flags to indicate which of the fourth 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 79 uM_fourth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
38% Activity at 39.5 uM_fourth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
39% Activity at 19.75 uM_fourth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
40% Activity at 9.875 uM_fourth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
41% Activity at 4.9375 uM_fourth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
42% Activity at 2.46875 uM_fourth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
43% Activity at 1.234375 uM_fourth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
44% Activity at 0.6171875 uM_fourth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
45% Activity at 0.3085938 uM_fourth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
46% Activity at 0.1542969 uM_fourth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
47IC50_Qualifier_1_Range1This qualifier is to be used with the next TID, IC50_1. If 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
48IC50_1_Range1IC50 value determined using sigmoidal dose response equationFloatμM
49Std.Err(IC50)_1_Range1Standard Error of IC50 valueFloatμM
50nH_1_Range1Hill coefficient determined using sigmoidal dose response equationFloat
51Excluded_Points_fifth_point_Range1Flags to indicate which of the fifth 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 79 uM_fifth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
53% Activity at 39.5 uM_fifth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
54% Activity at 19.75 uM_fifth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
55% Activity at 9.875 uM_fifth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
56% Activity at 4.9375 uM_fifth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
57% Activity at 2.46875 uM_fifth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
58% Activity at 1.234375 uM_fifth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
59% Activity at 0.6171875 uM_fifth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
60% Activity at 0.3085938 uM_fifth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
61% Activity at 0.1542969 uM_fifth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
62Excluded_Points_sixth_point_Range1Flags to indicate which of the sixth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
63% Activity at 79 uM_sixth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
64% Activity at 39.5 uM_sixth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
65% Activity at 19.75 uM_sixth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
66% Activity at 9.875 uM_sixth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
67% Activity at 4.9375 uM_sixth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
68% Activity at 2.46875 uM_sixth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
69% Activity at 1.234375 uM_sixth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
70% Activity at 0.6171875 uM_sixth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
71% Activity at 0.3085938 uM_sixth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
72% Activity at 0.1542969 uM_sixth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
73Excluded_Points_seventh_point_Range1Flags to indicate which of the seventh dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
74% Activity at 79 uM_seventh_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
75% Activity at 39.5 uM_seventh_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
76% Activity at 19.75 uM_seventh_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
77% Activity at 9.875 uM_seventh_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
78% Activity at 4.9375 uM_seventh_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
79% Activity at 2.46875 uM_seventh_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
80% Activity at 1.234375 uM_seventh_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
81% Activity at 0.6171875 uM_seventh_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
82% Activity at 0.3085938 uM_seventh_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
83% Activity at 0.1542969 uM_seventh_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
84Excluded_Points_eighth_point_Range1Flags to indicate which of the eighth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
85% Activity at 79 uM_eighth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
86% Activity at 39.5 uM_eighth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
87% Activity at 19.75 uM_eighth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
88% Activity at 9.875 uM_eighth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
89% Activity at 4.9375 uM_eighth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
90% Activity at 2.46875 uM_eighth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
91% Activity at 1.234375 uM_eighth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
92% Activity at 0.6171875 uM_eighth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
93% Activity at 0.3085938 uM_eighth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
94% Activity at 0.1542969 uM_eighth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
95IC50_Qualifier_2_Range1This qualifier is to be used with the next TID, IC50_2. If 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
96IC50_2_Range1IC50 value determined using sigmoidal dose response equationFloatμM
97Std.Err(IC50)_2_Range1Standard Error of IC50 valueFloatμM
98nH_2_Range1Hill coefficient determined using sigmoidal dose response equationFloat
99Excluded_Points_ninth_point_Range1Flags to indicate which of the ninth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
100% Activity at 79 uM_ninth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
101% Activity at 39.5 uM_ninth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
102% Activity at 19.75 uM_ninth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
103% Activity at 9.875 uM_ninth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
104% Activity at 4.9375 uM_ninth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
105% Activity at 2.46875 uM_ninth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
106% Activity at 1.234375 uM_ninth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
107% Activity at 0.6171875 uM_ninth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
108% Activity at 0.3085938 uM_ninth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
109% Activity at 0.1542969 uM_ninth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
110Excluded_Points_tenth_point_Range1Flags to indicate which of the tenth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
111% Activity at 79 uM_tenth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
112% Activity at 39.5 uM_tenth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
113% Activity at 19.75 uM_tenth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
114% Activity at 9.875 uM_tenth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
115% Activity at 4.9375 uM_tenth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
116% Activity at 2.46875 uM_tenth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
117% Activity at 1.234375 uM_tenth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
118% Activity at 0.6171875 uM_tenth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
119% Activity at 0.3085938 uM_tenth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
120% Activity at 0.1542969 uM_tenth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
121Excluded_Points_eleventh_point_Range1Flags to indicate which of the eleventh dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
122% Activity at 79 uM_eleventh_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
123% Activity at 39.5 uM_eleventh_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
124% Activity at 19.75 uM_eleventh_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
125% Activity at 9.875 uM_eleventh_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
126% Activity at 4.9375 uM_eleventh_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
127% Activity at 2.46875 uM_eleventh_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
128% Activity at 1.234375 uM_eleventh_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
129% Activity at 0.6171875 uM_eleventh_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
130% Activity at 0.3085938 uM_eleventh_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
131% Activity at 0.1542969 uM_eleventh_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
132Excluded_Points_twelfth_point_Range1Flags to indicate which of the twelfth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
133% Activity at 79 uM_twelfth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
134% Activity at 39.5 uM_twelfth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
135% Activity at 19.75 uM_twelfth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
136% Activity at 9.875 uM_twelfth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
137% Activity at 4.9375 uM_twelfth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
138% Activity at 2.46875 uM_twelfth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
139% Activity at 1.234375 uM_twelfth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
140% Activity at 0.6171875 uM_twelfth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
141% Activity at 0.3085938 uM_twelfth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
142% Activity at 0.1542969 uM_twelfth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
143IC50_Qualifier_3_Range1This qualifier is to be used with the next TID, IC50_3. If 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
144IC50_3_Range1IC50 value determined using sigmoidal dose response equationFloatμM
145Std.Err(IC50)_3_Range1Standard Error of IC50 valueFloatμM
146nH_3_Range1Hill coefficient determined using sigmoidal dose response equationFloat
147Excluded_Points_thirteenth_point_Range1Flags to indicate which of the thirteenth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
148% Activity at 79 uM_thirteenth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
149% Activity at 39.5 uM_thirteenth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
150% Activity at 19.75 uM_thirteenth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
151% Activity at 9.875 uM_thirteenth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
152% Activity at 4.9375 uM_thirteenth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
153% Activity at 2.46875 uM_thirteenth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
154% Activity at 1.234375 uM_thirteenth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
155% Activity at 0.6171875 uM_thirteenth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
156% Activity at 0.3085938 uM_thirteenth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
157% Activity at 0.1542969 uM_thirteenth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
158Excluded_Points_fourteenth_point_Range1Flags to indicate which of the fourteenth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
159% Activity at 79 uM_fourteenth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
160% Activity at 39.5 uM_fourteenth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
161% Activity at 19.75 uM_fourteenth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
162% Activity at 9.875 uM_fourteenth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
163% Activity at 4.9375 uM_fourteenth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
164% Activity at 2.46875 uM_fourteenth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
165% Activity at 1.234375 uM_fourteenth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
166% Activity at 0.6171875 uM_fourteenth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
167% Activity at 0.3085938 uM_fourteenth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
168% Activity at 0.1542969 uM_fourteenth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
169Excluded_Points_fifteenth_point_Range1Flags to indicate which of the fifteenth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
170% Activity at 79 uM_fifteenth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
171% Activity at 39.5 uM_fifteenth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
172% Activity at 19.75 uM_fifteenth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
173% Activity at 9.875 uM_fifteenth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
174% Activity at 4.9375 uM_fifteenth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
175% Activity at 2.46875 uM_fifteenth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
176% Activity at 1.234375 uM_fifteenth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
177% Activity at 0.6171875 uM_fifteenth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
178% Activity at 0.3085938 uM_fifteenth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
179% Activity at 0.1542969 uM_fifteenth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
180Excluded_Points_sixteenth_point_Range1Flags to indicate which of the sixteenth dose-response points were excluded from analysis. (1) means the titration point was excluded and (0) means the point was not excluded.String
181% Activity at 79 uM_sixteenth_point_Range1 (79μM**)% Inhibition at a given concentrationFloat%
182% Activity at 39.5 uM_sixteenth_point_Range1 (39.5μM**)% Inhibition at a given concentrationFloat%
183% Activity at 19.75 uM_sixteenth_point_Range1 (19.75μM**)% Inhibition at a given concentrationFloat%
184% Activity at 9.875 uM_sixteenth_point_Range1 (9.875μM**)% Inhibition at a given concentrationFloat%
185% Activity at 4.9375 uM_sixteenth_point_Range1 (4.9375μM**)% Inhibition at a given concentrationFloat%
186% Activity at 2.46875 uM_sixteenth_point_Range1 (2.46875μM**)% Inhibition at a given concentrationFloat%
187% Activity at 1.234375 uM_sixteenth_point_Range1 (1.23438μM**)% Inhibition at a given concentrationFloat%
188% Activity at 0.6171875 uM_sixteenth_point_Range1 (0.617188μM**)% Inhibition at a given concentrationFloat%
189% Activity at 0.3085938 uM_sixteenth_point_Range1 (0.308594μM**)% Inhibition at a given concentrationFloat%
190% Activity at 0.1542969 uM_sixteenth_point_Range1 (0.154297μM**)% Inhibition at a given concentrationFloat%
191IC50_Qualifier_4_Range1This qualifier is to be used with the next TID, IC50_4. If 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
192IC50_4_Range1IC50 value determined using sigmoidal dose response equationFloatμM
193Std.Err(IC50)_4_Range1Standard Error of IC50 valueFloatμM
194nH_4_Range1Hill coefficient determined using sigmoidal dose response equationFloat
* Activity Concentration. ** Test Concentration.
Additional Information
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Grant Number: 1R21NS059422-01

Data Table (Concise)
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