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BioAssay: AID 540309

Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 3 (S1P3)

Name: Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 3 (S1P3). ..more
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 Tested Compounds
 Tested Compounds
All(5)
 
 
Probe(5)
 
 
Active(5)
 
 
 Tested Substances
 Tested Substances
All(5)
 
 
Probe(5)
 
 
Active(5)
 
 
AID: 540309
Data Source: The Scripps Research Institute Molecular Screening Center (S1P3_AG_SUMMARY)
BioAssay Type: Panel, Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2011-07-27
Modify Date: 2012-03-02

Data Table ( Complete ):           View Probe Data    View Active Data    View All Data
Target
BioActive Compounds: Chemical Probe: 5    Active: 5
Related Experiments
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AIDNameTypeComment
373S1P3 Agonist Primary HTS and Confirmation AssaysScreeningdepositor-specified cross reference: Primary screen (S1P3 agonists in singlicate)
439S1P3 Agonist Dose-Response Potency AssayConfirmatorydepositor-specified cross reference: Dose response (S1P3 agonists in triplicate)
1192Dose Response Cell-Based Assay for Agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Purchased AnaloguesConfirmatorydepositor-specified cross reference: Dose response (S1P3 agonists in triplicate)
540344Late-stage assay provider results from the probe development effort to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): luminescence-based cell-based dose response counterscreen assay to determine cytotoxicity of agonist compoundsConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (Cytotoxicity with 6 replicates)
540349Late-stage fluorescence-based cell-based dose response assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Synthesized compoundsConfirmatorydepositor-specified cross reference: Late stage dose response (S1P3 agonists in triplicate)
540351Late-stage counterscreen panel assay for S1P3 agonists: Ricerca HitProfilingScreen + CYP450Otherdepositor-specified cross reference: Late stage counterscreen (Ricerca HitProfilingScreen + CYP450)
540366Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 4 (S1P4) agonist assayConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (S1P4 agonists in triplicate)
540367Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 2 (S1P2) agonist assayConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (S1P2 agonists in triplicate)
540368Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 1 (S1P1) agonist assay Set 3Confirmatorydepositor-specified cross reference: Late stage dose response counterscreen (S1P1 agonists in triplicate)
540369Late-stage fluorescence-based dose-response cell-based counterscreen assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): Sphingosine 1-Phosphate Receptor 5 (S1P5) agonist assayConfirmatorydepositor-specified cross reference: Late stage dose response counterscreen (S1P5 agonists with six replicates)
588327Late-stage assay to identify agonists of the Sphingosine 1-Phosphate Receptor 3 (S1P3): radiometric-based cell-based dose response S1P agonist competition binding assayConfirmatorydepositor-specified cross reference: Late-stage dose response (S1P agonist competition binding in triplicate)
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRISMC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Germana Sanna, TSRI
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number: 1R03MH076534-01
Grant Proposal PI: Germana Sanna, TSRI
External Assay ID: S1P3_AG_SUMMARY

Name: Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 3 (S1P3).

Description:

Sphingosine 1-phosphate (S1P) influences heart rate (1-2), coronary artery caliber, endothelial integrity, lung epithelial integrity and lymphocyte recirculation (2-5) through five related high affinity G-protein coupled receptors (6). Inhibition of lymphocyte recirculation by nonselective S1P receptor agonists produces clinical immunosuppression preventing transplant rejection, but is associated with transient bradycardia. Understanding the contribution of individual receptors has been limited in the past by the unavailability of selective agonists or antagonists for the five receptor subtypes. S1P3 receptor subtype plays a critical role in cardiac rhythm and lung epithelial barrier function. S1P1 and S1P3 are coexpressed in some cells, especially endothelium. In lung, S1P3 is exclusively expressed upon pulmonary epithelium and activation of S1P3 results in rapid dissolution of epithelial tight junctions, measured by both ZO-1 and claudin degradation (7). This produces acute development of paracellular gaps and pulmonary edema that is reversed in the S1P3-deltant mouse. S1P3 antagonists would be of great utility in the assessment of mechanisms of Adult Respiratory Distress Syndrome (ARDS). The use of SEW2871 together with the S1P3-deletant mice shows that S1P1 and S1P3 appear to have mutually exclusive roles: activation of S1P1 is sufficient to control lymphocyte numbers and plays no discernable role in control of sinus rhythm, whereas S1P3 regulates sinus rhythm and not lymphocyte recirculation. Agonists and antagonists of S1P3 may be useful probes of cardiac function in vivo. Understanding the role(s) of S1P receptor subtypes in the control of endothelial and epithelial integrity will allow the identification of selective immunosuppressive S1P3 receptor modulators that could be of use in the control of cardiac function and the prevention of ARDS. S1P receptor subtype selective agonists and antagonists will be of broad utility in understanding cell functions in vitro and vascular physiology in vivo.

Summary of Probe Development Effort:

Following primary HTS in singlicate (AID 373) and titration assays in triplicate to determine potency (AID 439 and AID 1192), certain compounds were identified as possible candidates for probe development. A total of 65 structures were clustered using a 0.7 similarity threshold and Leadscope fingerprints to identify 13 clusters and 19 singletons. The top 10 clusters and singletons were identified based on their best activity. Structural classes that showed activity against S1P1 were removed. Four of these compounds, SID 112379033, SID 112379041, SID 112379040, and SID 112379039 demonstrated an improvement over prior art, and therefore constitute novel S1P3 agonist probes.

All AIDs that contain results associated with this project can be found in the "Related Bioassays" section of this Summary AID. A probe report has been submitted. The results of our probe development efforts can be found at http://mlpcn.florida.scripps.edu/index.php/probes/probe-reports. A probe report for SID 112379033, SID 112379041, SID 112379040, and SID 112379039 can be found in the Molecular Libraries Bookshelf (PubMed Books) (http://www.ncbi.nlm.nih.gov/books) under ML003, ML004, ML005, and ML006, respectively.

References:

1. Forrest, M., et al., Immune cell regulation and cardiovascular effects of sphingosine 1-phosphate receptor agonists in rodents are mediated via distinct receptor subtypes. J Pharmacol Exp Ther, 2004. 309(2): p. 758-68.
2. Sanna, M.G., et al., Sphingosine 1-phosphate (S1P) receptor subtypes S1P1 and S1P3, respectively, regulate lymphocyte recirculation and heart rate. J Biol Chem, 2004. 279(14): p. 13839-48.
3. Alfonso, C., M.G. McHeyzer-Williams, and H. Rosen, CD69 down-modulation and inhibition of thymic egress by short- and long-term selective chemical agonism of sphingosine 1-phosphate receptors. Eur J Immunol, 2006. 36(1): p. 149-59.
4. Jo, E., et al., S1P1-selective in vivo-active agonists from high-throughput screening: off-the-shelf chemical probes of receptor interactions, signaling, and fate. Chem Biol, 2005. 12(6): p. 703-15.
5. Wei, S.H., et al., Sphingosine 1-phosphate type 1 receptor agonism inhibits transendothelial migration of medullary T cells to lymphatic sinuses. Nat Immunol, 2005. 6(12): p. 1228-35.
6. Mandala, S., et al., Alteration of lymphocyte trafficking by sphingosine-1-phosphate receptor agonists. Science, 2002. 296(5566): p. 346-9.
7. Goetzl, E.J. and H. Rosen, Regulation of immunity by lysosphingolipids and their G protein-coupled receptors. J Clin Invest, 2004. 114(11): p. 1531-7.

Keywords:

summary, summary AID, sphingosine receptor, sphingosine-1-phosphate receptor 3, S1P3, EDG-3, endothelial differentiation gene-3, G protein-coupled receptor, GPCR, agonist, activator, Adult Respiratory Distress Syndrome, ARDS, probe, probe report, ML003, ML004, ML005, ML006, Scripps, Scripps Florida, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Bookshelf, Molecular Library Probe Production Centers Network, MLPCN
Panel Information
Related BioAssays
    Data Table(Active)    Data Table(All)Show more
PID§NameSubstancePanel TargetsDescriptionAdditional Information
ActiveInactive
1AID 540349_1sphingosine 1-phosphate receptor 3 [Homo sapiens] [gi:38788193]
Taxonomy id: 9606
Gene id: 1903
2AID 540349_2sphingosine 1-phosphate receptor 3 [Homo sapiens] [gi:38788193]
Taxonomy id: 9606
Gene id: 1903
3AID 540349_3sphingosine 1-phosphate receptor 3 [Homo sapiens] [gi:38788193]
Taxonomy id: 9606
Gene id: 1903
4AID 540344
5AID 540369Sphingosine-1-phosphate receptor 5 [Homo sapiens] [gi:21961528]
Taxonomy id: 9606
Gene id: 53637
6AID 540366Sphingosine-1-phosphate receptor 4 [Homo sapiens] [gi:15929025]
Taxonomy id: 9606
Gene id: 8698
7AID 540367sphingosine 1-phosphate receptor 2 [Homo sapiens] [gi:134244587]
Taxonomy id: 9606
Gene id: 9294

§ Panel component ID.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML249
PubChem Substance ID (SID) for probe 1: 124360653
PubChem Compound ID (CID) for probe 1: 17253208
Probe type for probe 1: Agonist
IC50/EC50 (nM) for probe 1: <132
Target for probe 1: S1P3 (gi: 119583162)
Disease relevance for probe 1: Hypertension
Anti-target for probe 1: S1P1
Fold selectivity for probe 1: >213
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK133421/ (ID: 3025156)
Grant number for probe 1: MH076534-01
PubMed Publication ID (PMID) for probe 1: 22971058,24135724
NCBI Book chapter title for probe 1: Probe Development Efforts for an Allosteric Agonist of the Sphingosine 1-phosphate Receptor 3 (S1P3)
Result Definitions
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TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1ML #Unique alphanumeric identifier assigned to a chemical probe molecule within the Molecular Libraries Probe Production Centers Network (MLPCN).String
2EC50 [AID 540349_1]*Data from AID 540349_1. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.1sphingosine 1-phosphate receptor 3 [Homo sapiens]FloatμM
3EC50 [AID 540349_2]*Data from AID 540349_2. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.2sphingosine 1-phosphate receptor 3 [Homo sapiens]FloatμM
4EC50 [AID 540349_3]*Data from AID 540349_3. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.3sphingosine 1-phosphate receptor 3 [Homo sapiens]FloatμM
5Qualifier [AID 540344]Data from AID 504344. Activity Qualifier identifies if the resultant data CC50 came from a fitted curve or was determined manually to be less than or greater than its listed CC50 concentration.4String
6CC50 [AID 540344]*Data from AID 540344. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.4FloatμM
7Qualifier [AID 540369]Data from AID 504369. Activity Qualifier identifies if the resultant data EC50 came from a fitted curve or was determined manually to be less than or greater than its listed EC50 concentration.5Sphingosine-1-phosphate receptor 5 [Homo sapiens]String
8EC50 [AID 540369]*Data from AID 540369. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.5FloatμM
9Qualifier [AID 540366]Data from AID 504366. Activity Qualifier identifies if the resultant data EC50 came from a fitted curve or was determined manually to be less than or greater than its listed EC50 concentration.6Sphingosine-1-phosphate receptor 4 [Homo sapiens]String
10EC50 [AID 540366]*Data from AID 540366. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.6FloatμM
11Qualifier [AID 540367]Data from AID 504367. Activity Qualifier identifies if the resultant data EC50 came from a fitted curve or was determined manually to be less than or greater than its listed EC50 concentration.7sphingosine 1-phosphate receptor 2 [Homo sapiens]String
12EC50 [AID 540367]*Data from AID 540367. The value for concentration at which 50% inhibition is observed; EC50 shown in uM.7FloatμM

* Activity Concentration. § Panel component ID.
Additional Information
Grant Number: 1R03MH076534-01

Classification
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