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BioAssay: AID 2449

PPAR gamma Selectivity Assay For Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2

Thyroid receptor (TR) regulates many homeostatic processes including basal metabolism, cardiovascular function, body weight, and lipid trafficking. TR modulators are potential therapeutics for obesity and hyperlipidemias but current thyroid analogs have undesirable side effects, particularly cardiac stimulation. This secondary assay characterized compounds identified in the qHTS for Inhibitors of more ..
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 Tested Compounds
 Tested Compounds
All(25)
 
 
Active(4)
 
 
Inactive(19)
 
 
Inconclusive(2)
 
 
 Tested Substances
 Tested Substances
All(25)
 
 
Active(4)
 
 
Inactive(19)
 
 
Inconclusive(2)
 
 
AID: 2449
Data Source: NCGC (TR541)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-03-02
Hold-until Date: 2010-06-15
Modify Date: 2010-06-15

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 4
Depositor Specified Assays
AIDNameTypeProbeComment
1469qHTS for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2confirmatory
1479Total Fluorescence Counterscreen for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2confirmatory
1570Concentration Response Confirmation Assay for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2, Fluorescein Fluoroprobeconfirmatory
1571Total Fluorescence Confirmation Counterscreen for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2, Fluorescein Fluoroprobeconfirmatory
1572Total Fluorescence Confirmation Counterscreen for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2confirmatory
1573Concentration Response Confirmation Assay for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2confirmatory
1485Quantitative High-Throughput Screen for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2: Summarysummary
2512qHTS Assay for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2: Probe Summarysummary1
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: DK058080
Assay Provider: R. Kip Guy, St. Jude Children's Research Hospital

NCGC Assay Overview:

Thyroid receptor (TR) regulates many homeostatic processes including basal metabolism, cardiovascular function, body weight, and lipid trafficking. TR modulators are potential therapeutics for obesity and hyperlipidemias but current thyroid analogs have undesirable side effects, particularly cardiac stimulation. This secondary assay characterized compounds identified in the qHTS for Inhibitors of the Interaction of Thyroid Hormone Receptor and Steroid Receptor Coregulator 2 (PubChem AID 1485) for selectivity against PPAR gamma. This assay detects interaction of the ligand-binding domain of human PPAR gamma with a Texas Red labeled DRIP-2 peptide, corresponding to a 20 amino acid region of the nuclear receptor interaction domain (Feau et al., 2009). Small molecule inhibitors that block the interaction of PPAR and DRIP-2 are detected by a decrease in fluorescence polarization.

Feau C, Arnold LA, Kosinski A, Zhu F, Connelly M, Guy RK. Novel flufenamic acid analogues as inhibitors of androgen receptor mediated transcription.
ACS Chem Biol. 2009 4(10):834-43.
Protocol
NCGC Assay Protocol Summary:

Twenty uL/well 2 uM PPAR gamma and 10 nM Tx-DRIP-2 peptide in protein buffer (20 mM TRIS, pH 7.5, 100 mM NaCl, 0.01% NP-40, 20 uM roziglitazone, and 4% DMSO) was dispensed into black solid 384-well plates (Costar 3710) using a Biomek FX (Beckman Coulter) liquid handling system. Following transfer of 260 nL compound (ranging from 7 nM to 130 uM), the plates were incubated 3 hr at ambient temperature. The plates were read by an Envision (Perkin Elmer) to detect Texas Red polarization of Tx-DRIP-2 (555 nm excitation and 632 nm emission).
Comment
Compound Ranking:

Active compounds were assigned a score between 50 and 100 based on compound potency. Inactive compounds were assigned a score of 0 and inconclusive compounds were assigned a score of 25.
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1ActivityIndicates type of activity observed: active or inactive.String
2Potency*The concentration of sample yielding half-maximal inhibition.FloatμM
3Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration.
Additional Information
Grant Number: DK058080

Data Table (Concise)
Classification
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