Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 4 (S1P4)
Name: Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 4 (S1P4) ..more
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Michael Oldstone, TSRI
Network: Molecular Library Probe Production Center Network (MLPCN)
Grant Proposal Number: U01 AI074564 Fast Track
Grant Proposal PI: Michael Oldstone, TSRI
External Assay ID: S1P4_AG_LEADS_SUMMARY
Name: Summary of probe development efforts to identify agonists of Sphingosine 1-Phosphate Receptor 4 (S1P4)
Pandemic influenza represents a significant public health threat, due in part to immune cell-mediated lung tissue damage induced during viral infection. Sphingosine 1-phosphate (S1P) is a bioactive phospholipid released by activated blood platelets and serves to influence endothelial integrity, lung epithelial integrity (1), and lymphocyte recirculation (2-5) through five related high affinity G-protein coupled receptors. Recently, modulation of S1P receptors locally in the lungs was shown to alter dendritic cell activation and accumulation in the mediastinal lymph nodes, resulting in blunted T cell responses and control of immunopathological features of influenza virus infection (6). Reports showing that S1P5 expression is very low in dendritic cells but that S1P4 is highly expressed (7), suggest that chemical activation of the S1P4 receptor subtype in the airways could be efficient at controlling the immunopathological response to viral infection. S1P4 is coupled to Ga(i) and Ga(o) G-proteins and activates ERK MAPK and PLC downstream pathways (8). Thus, the identification of compounds that act as selective S1P4 agonists will provide insight into S1P4 biology and may serve as useful tools to limit lung tissue injury resulting from influenza infection.
Summary of Probe Development Effort:
This probe development effort is focused on the identification of S1P4 agonists. Following uHTS primary, confirmation, and dose response screening and counterscreening against the S1P1 receptor, a number of compounds of interest emerged. SAR analysis by purchase and synthesis was conducted on the lead hit. Titration assays were performed to determine potency and selectivity against the other S1P family member receptors S1P1, S1P2, S1P3, and S1P5, as well as cytotoxicity. Compound SID 87544119, with an EC50 for S1P4 of 46.3 nM and EC50s > 50 uM for the other S1P family of receptors, was selected as an exemplar of the potent set of S1P4 agonists. Compound SID 87544119 is also inhibited by an S1P4 receptor-selective antagonist with an IC50 of 0.83 uM.
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 87544119 can be found in the Molecular Libraries Bookshelf (PubMed Books) (http://www.ncbi.nlm.nih.gov/books) under ML178.
1. Sanna, M.G., J. Liao, E. Jo, C. Alfonso, M.Y. Ahn, M.S. Peterson, B. Webb, S. Lefebvre, J. Chun, N. Gray, and H. Rosen, 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.
2. Forrest, M., S.Y. Sun, R. Hajdu, J. Bergstrom, D. Card, G. Doherty, J. Hale, C. Keohane, C. Meyers, J. Milligan, S. Mills, N. Nomura, H. Rosen, M. Rosenbach, G.J. Shei, Singer, II, M. Tian, S. West, V. White, J. Xie, R.L. Proia, and S. Mandala, 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.
3. Gon, Y., M.R. Wood, W.B. Kiosses, E. Jo, M.G. Sanna, J. Chun, and H. Rosen, S1P3 receptor-induced reorganization of epithelial tight junctions compromises lung barrier integrity and is potentiated by TNF. Proc Natl Acad Sci U S A, 2005. 102(26): p. 9270-5.
4. Wei, S.H., H. Rosen, M.P. Matheu, M.G. Sanna, S.K. Wang, E. Jo, C.H. Wong, I. Parker, and M.D. Cahalan, 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.
5. 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.
6. Jo, E., M.G. Sanna, P.J. Gonzalez-Cabrera, S. Thangada, G. Tigyi, D.A. Osborne, T. Hla, A.L. Parrill, and H. Rosen, 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.
7. Maeda, Y., Matsuyuki, H., Shimano, K., Kataoka, H., Sugahara, K., and Chiba, K., Migration of CD4 T cells and dendritic cells toward sphingosine 1-phosphate (S1P) is mediated by different receptor subtypes: S1P regulates the functions of murine mature dendritic cells via S1P receptor type 3. J Immunol, 2007. 178(6): p. 3437-46.
8. Toman, R.E. and S. Spiegel, Lysophospholipid receptors in the nervous system. Neurochem Res, 2002. 27(7-8): p. 619-27.
Summary AID, Sphingosine Receptor, Sphingosine-1-phosphate receptor 4, S1P4, S1PR4, EDG6, LPC1, agonist, activator, HTS, GPCR, 1536, Tango, FRET, GAL4-VP16, beta-arrestin, beta-lactamase, BLA, reporter gene, fluorescence, dose response, counterscreen, S1P1, EDG1, EDG-1, Scripps, Scripps Florida, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Library Probe Production Center Network, MLPCN.
§ Panel component ID.
Please see AIDs 1509, 1523, 1563 1686, 1701 for protocols performed in this probe development effort.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 2
MLP Probe ML# for probe 1: ML178
PubChem Substance ID (SID) for probe 1: 87544119
PubChem Compound ID (CID) for probe 1: 44620892
Probe type for probe 1: Agonist
IC50/EC50 (nM) for probe 1: 46.3
Target for probe 1: S1P4 (gi: 15929025)
Disease relevance for probe 1: influenza
Anti-target for probe 1: S1P1
Fold selectivity for probe 1: >1000
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK98915/ (ID: 2961648)
Grant number for probe 1: AI074564
MLP Probe ML# for probe 2: ML248
PubChem Substance ID (SID) for probe 2: 103911214
PubChem Compound ID (CID) for probe 2: 49835928
Probe type for probe 2: Agonist
IC50/EC50 (nM) for probe 2: <80
Target for probe 2: S1P4 (gi: 15929025)
Anti-target for probe 2: S1P1
Fold selectivity for probe 2: >316
NCBI Book chapter link for probe 2: http://www.ncbi.nlm.nih.gov/books/NBK143556/ (ID: 3036516)
Grant number for probe 2: AI074564
NCBI Book chapter title for probe 1: Probe Development Efforts to Identify Novel Agonists of the Sphingosine 1-phosphate Receptor 4 (S1P4)
NCBI Book chapter title for probe 2: Identification of a Novel Agonist of the Sphingosine 1-phosphate Receptor 4 (S1P4)
* Activity Concentration. § Panel component ID.