Bookmark and Share
BioAssay: AID 504660

Allosteric Agonists of the Human D1 Dopamine Receptor: qHTS

Dopamine receptors (DARs) are involved in the etiology and/or therapy of a number of neuropsychiatric and endocrine disorders. For instance, all receptor-based antiparkinsonian drugs work via stimulating the D1 DAR subtype whereas all FDA-approved antipsychotic agents are antagonists of this receptor. Most drugs targeting the D1 DAR are problematic, however, either being less efficacious as more ..
_
   
 Tested Compounds
 Tested Compounds
All(361338)
 
 
Active(3713)
 
 
Inactive(357664)
 
 
 Tested Substances
 Tested Substances
All(364628)
 
 
Active(3724)
 
 
Inactive(360904)
 
 
AID: 504660
Data Source: NCGC (DOP1001)
BioAssay Type: Primary, Primary Screening, Single Concentration Activity Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-04-13

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 3713
Related Experiments
AIDNameTypeComment
488989HTS Assay for Allosteric Agonists of the Human D1 Dopamine Receptor: SummarySummarydepositor-specified cross reference
488981HTS Assay for Allosteric Agonists of the Human D1 Dopamine Receptor: Primary Screen for AgonistsConfirmatorysame project related to Summary assay
488982HTS Assay for Allosteric Agonists of the Human D1 Dopamine Receptor: Primary Screen for PotentiatorsConfirmatorysame project related to Summary assay
488983HTS Assay for Allosteric Agonists of the Human D1 Dopamine Receptor: Primary Screen for AntagonistsConfirmatorysame project related to Summary assay
Description:
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: NS064831-01
Assay Submitter (PI): David Sibley

Dopamine receptors (DARs) are involved in the etiology and/or therapy of a number of neuropsychiatric and endocrine disorders. For instance, all receptor-based antiparkinsonian drugs work via stimulating the D1 DAR subtype whereas all FDA-approved antipsychotic agents are antagonists of this receptor. Most drugs targeting the D1 DAR are problematic, however, either being less efficacious as desired or possessing limiting side effects, most of which are due to reactivity at other receptors. One approach towards improved receptor specificity is to identify allosteric ligands that bind to less conserved regions of the receptor and therefore have the potential to be much more selective. The goal of this project is to use high throughput screening approaches to identify and develop novel, highly selective small molecule allosteric modulators of the D1 DAR for use as in vitro and in vivo pharmacological tools and in proof-of-concept experiments in animal models of neuropsychiatric disease. There are three different types of allosteric modulators that we are seeking, two of which will stimulate or augment receptor signaling, allosteric agonists and potentiators, while the third, allosteric antagonists, will attenuate receptor signaling. The present primary screening assay measures compound agonism by tracking calcium flux in a force-coupled, inducible Hek293 Trex D1 cell line.
Protocol
Freshly passaged cells are plated in 1536 well black, clear bottom plates at a density of 4000 cells/well in 3 ul of complete media containing 1x tetracycline inducer. After an overnight incubation at 30 C in 5% CO2, cells are loaded with 2 ul of the no wash Calcium Assay Kit (ABD Bioquest) and incubated at room temperature for 30 to 90 minutes. Agonist read: A 10 cycle (1 second/cycle) baseline measurement is taken and then the FDSS online 1536 pintool delivers 23 nl of compound library (as well as dopamine controls) to the plate. The plate is measured for kinetically at 1 cycle/sec for 180 seconds and is considered to be the agonist portion of the assay (eg, compounds which activate a positive calcium response during this portion are flagged as potential agonists). Following the 180 second read, 1 ul of either an EC20 (positive modulation assay) or EC80 (negative modulation assay) is delivered by an onboard pipette head. Measurements are taken kinetically for an additional 180 seconds.
Comment
Compound Ranking:
For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds with more than thirty percent activity, activity score was given as 50.
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Activity at 2.0 uM (2μM**)% Activity at given concentration.Float%
2Compound QCOriginator of sample that provided QC validation.String

** Test Concentration.
Additional Information
Grant Number: NS064831-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
PageFrom: