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BioAssay: AID 1401

Quantitative High-Throughput Screen for Agonists of Thyroid Stimulating Hormone Receptor: Summary

TSH is an alpha/beta heterodimeric glycoprotein hormone secreted from the anterior pituitary gland which belongs to the glycoprotein hormone family. The actions of TSH are mediated by a seven-transmembrane receptor, which upon TSH binding couples preferentially to the G-alpha (s) protein (Gs), resulting in activation of adenylate cyclase and increase in cyclic adenosine 3', 5' monophosphate more ..
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 Tested Compounds
 Tested Compounds
All(126)
 
 
Probe(2)
 
 
Active(33)
 
 
Inactive(64)
 
 
Inconclusive(29)
 
 
 Tested Substances
 Tested Substances
All(126)
 
 
Probe(2)
 
 
Active(33)
 
 
Inactive(64)
 
 
Inconclusive(29)
 
 
AID: 1401
Data Source: NCGC (TSHR_summary)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Screening Center Network
BioAssay Version:
Deposit Date: 2008-09-29
Modify Date: 2009-11-06

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: Chemical Probe: 2    Active: 33
Related Experiments
AIDNameTypeComment
926qHTS Assay for Agonists of the Thyroid Stimulating Hormone ReceptorConfirmatorydepositor-specified cross reference
933Secondary Concentration-Response Assay for Agonists of the Thyroid Stimulating Hormone Receptor: HTRF Activity DetectionConfirmatorydepositor-specified cross reference
938qHTS Assay for Agonists of the Thyroid Stimulating Hormone Receptor: Activators of Intracellular cAMP Concentrations in Parental HEK 293Confirmatorydepositor-specified cross reference
939Confirmation Concentration-Response Assay for Agonists of the Thyroid Stimulating Hormone ReceptorConfirmatorydepositor-specified cross reference
953Confirmation Concentration-Response Assay for Agonists of the Thyroid Stimulating Hormone Receptor: Activators of Intracellular cAMP Concentrations in Parental HEK 293Confirmatorydepositor-specified cross reference
1402Counterscreen for Agonists of Thyroid Stimulating Hormone Receptor: HTRF Activity in a Luteinizing Hormone Receptor Cell LineConfirmatorydepositor-specified cross reference
1403Counterscreen for Agonists of Thyroid Stimulating Hormone Receptor: HTRF Activity in a Follicle Stimulating Hormone Receptor Cell LineConfirmatorydepositor-specified cross reference
2104Secondary Concentration-Response Assay for Agonists of the Thyroid Stimulating Hormone Receptor: ELISA Activity DetectionConfirmatorydepositor-specified cross reference
Description:
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Screening Centers Network [MLSCN]
MLSCN Grant: 1 X01 MH080680-01
PI Name: Dr. Marvin Gershengorn, NIH

NCGC Assay Overview:

Thyroid Stimulating Hormone Receptor

TSH is an alpha/beta heterodimeric glycoprotein hormone secreted from the anterior pituitary gland which belongs to the glycoprotein hormone family. The actions of TSH are mediated by a seven-transmembrane receptor, which upon TSH binding couples preferentially to the G-alpha (s) protein (Gs), resulting in activation of adenylate cyclase and increase in cyclic adenosine 3', 5' monophosphate (cAMP). The TSH receptor (THSR) is mainly expressed in thyroid follicular cells and regulates their growth and function. Recombinant TSH is used to activate TSHR in patients with thyroid cancer receiving thyroid hormone suppression therapy and to screen for residual tumor after surgery, but it is expensive and must be administered intramuscularly. An orally active small molecule TSHR agonist would serve as an invaluable research tool for studying TSHR pharmacology and physiology, and would have multiple advantages in therapeutic settings. However, no selective small molecule agonist of the TSH receptor exists, and no small molecule screen for TSHR agonists has ever been reported.
The primary assay utilized a cyclic nucleotide gated ion channel (CNG) as a biosensor for cAMP induction by potential TSHR agonists. A confirmatory assay of putative TSHR agonists was conducted on a different TSHR transfected HEK 293 cell line, using an orthogonal detection technology to measure cAMP stimulation: antibody-based Homogeneous Time Resolved Fluorescence (HTRF). In order to confirm specificity of the compounds to the TSHR, similar family members were also assayed including the Follicle Stimulating Hormone (FSH) receptor and Luteinizing Hormone (LH) receptor using transfected HEK 293 cell lines and measured using the HTRF format.
Protocol
Please refer to other AIDs (926, 938, 939, 953, 933, 1402, 1403, 2104) for detailed assay protocols.
Comment
For the purposes of summarizing the probe activities from the project, MLSCN probes were given a score of 100. Molecules in the prior art were given a score of 80. Other, less active molecules in the same chemical series as the probe molecules were given a score of 50. Inactive analogues from these series were given a score of 0.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: HEK293
From MLP Probe Report:
Probe count: 2
MLP Probe ML# for probe 1: ML067
PubChem Substance ID (SID) for probe 1: 3714076
PubChem Compound ID (CID) for probe 1: 2998659
Probe type for probe 1: Agonist
IC50/EC50 (nM) for probe 1: 11500
Anti-target for probe 1: FSHR, LHCGR
Fold selectivity for probe 1: Inactive @100 uM
Grant number for probe 1: MH080680-01
MLP Probe ML# for probe 2: ML109
PubChem Substance ID (SID) for probe 2: 26755506
PubChem Compound ID (CID) for probe 2: 25246343
Probe type for probe 2: Agonist
IC50/EC50 (nM) for probe 2: 7943
Target for probe 2: TSH receptor (gi: 38016895)
Disease relevance for probe 2: Inflammation and cancer
Anti-target for probe 2: FSHR, LHR
Fold selectivity for probe 2: >10
NCBI Book chapter link for probe 2: http://www.ncbi.nlm.nih.gov/books/NBK56234/ (ID: 2509788)
Grant number for probe 2: MH080680-01
PubMed Publication ID (PMID) for probe 1: 18216391
PubMed Publication ID (PMID) for probe 2: 19592511
NCBI Book chapter title for probe 2: Identification of Potent and Selective Thyroid Stimulating Hormone Receptor Agonists
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Thyroid Stimulating Hormone Receptor PotencyThe concentration of sample yielding half-maximal activation of the thyroid stimulating hormone receptor.FloatμM
2Thyroid Stimulating Hormone Receptor EfficacyThe maximal activation of the thyroid stimulating hormone receptor.Float%
3Thyroid Stimulating Hormone HTRFWhether a concentration-response was observed for the sample in a secondary assay of TSH receptor activity.String
4Luteinizing Hormone ReceptorWhether a concentration-response was observed for the sample in a secondary assay of LH receptor activity.String
5Follicle Stimulating Hormone ReceptorWhether a concentration-response was observed for the sample in a secondary assay of FSH receptor activity.String
Additional Information
Grant Number: X01 MH080680-01

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