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
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).
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.
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.
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§ 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)
* Activity Concentration. § Panel component ID.