{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T15:51:12Z","timestamp":1791474672720,"version":"4.3.4"},"reference-count":44,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2020,9,4]],"date-time":"2020-09-04T00:00:00Z","timestamp":1599177600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Schweizerischer Nationalfonds zur F\u00f6rderung der Wissenschaftlichen Forschung","doi-asserted-by":"publisher","award":["310030_135619"],"award-info":[{"award-number":["310030_135619"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["GRK2158, INST 208\/690-1"],"award-info":[{"award-number":["GRK2158, INST 208\/690-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000921","name":"European Cooperation in Science and Technology","doi-asserted-by":"publisher","award":["CA15135 and CA18240"],"award-info":[{"award-number":["CA15135 and CA18240"]}],"id":[{"id":"10.13039\/501100000921","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Multiple sclerosis (MS) is a chronic, inflammatory, autoimmune disease of the central nervous system (CNS) which is associated with lower life expectancy and disability. The experimental antigen-induced encephalomyelitis (EAE) in mice is a useful animal model of MS, which allows exploring the etiopathogenetic mechanisms and testing novel potential therapeutic drugs. A new therapeutic paradigm for the treatment of MS was introduced in 2010 through the sphingosine 1-phosphate (S1P) analogue fingolimod (FTY720, Gilenya\u00ae), which acts as a functional S1P1 antagonist on T lymphocytes to deplete these cells from the blood. In this study, we synthesized two novel structures, ST-1893 and ST-1894, which are derived from fingolimod and chemically feature a morpholine ring in the polar head group. These compounds showed a selective S1P1 activation profile and a sustained S1P1 internalization in cultures of S1P1-overexpressing Chinese hamster ovary (CHO)-K1 cells, consistent with a functional antagonism. In vivo, both compounds induced a profound lymphopenia in mice. Finally, these substances showed efficacy in the EAE model, where they reduced clinical symptoms of the disease, and, on the molecular level, they reduced the T-cell infiltration and several inflammatory mediators in the brain and spinal cord. In summary, these data suggest that S1P1-selective compounds may have an advantage over fingolimod and siponimod, not only in MS but also in other autoimmune diseases.<\/jats:p>","DOI":"10.3390\/ijms21186463","type":"journal-article","created":{"date-parts":[[2020,9,4]],"date-time":"2020-09-04T11:24:24Z","timestamp":1599218664000},"page":"6463","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Morpholino Analogues of Fingolimod as Novel and Selective S1P1 Ligands with In Vivo Efficacy in a Mouse Model of Experimental Antigen-Induced Encephalomyelitis"],"prefix":"10.3390","volume":"21","author":[{"given":"Bisera","family":"Stepanovska","sequence":"first","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Inselspital INO-F, CH-3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandra","family":"Zivkovic","sequence":"additional","affiliation":[{"name":"Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University D\u00fcsseldorf, Universitaetsstr. 1, D-40225 Duesseldorf, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaby","family":"Enzmann","sequence":"additional","affiliation":[{"name":"Theodor Kocher Institute, University of Bern, Freiestrasse 1, CH-3012 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Tietz","sequence":"additional","affiliation":[{"name":"Theodor Kocher Institute, University of Bern, Freiestrasse 1, CH-3012 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Homann","sequence":"additional","affiliation":[{"name":"Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert Allee 114\u2013116, D-14558 Nuthetal, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1888-9595","authenticated-orcid":false,"given":"Burkhard","family":"Kleuser","sequence":"additional","affiliation":[{"name":"Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert Allee 114\u2013116, D-14558 Nuthetal, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Britta","family":"Engelhardt","sequence":"additional","affiliation":[{"name":"Theodor Kocher Institute, University of Bern, Freiestrasse 1, CH-3012 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3336-1710","authenticated-orcid":false,"given":"Holger","family":"Stark","sequence":"additional","affiliation":[{"name":"Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University D\u00fcsseldorf, Universitaetsstr. 1, D-40225 Duesseldorf, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1615-5691","authenticated-orcid":false,"given":"Andrea","family":"Huwiler","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Inselspital INO-F, CH-3010 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2286","DOI":"10.1089\/ars.2012.5068","article-title":"Multiple sclerosis: Molecular mechanisms and therapeutic opportunities","volume":"19","author":"Miljkovic","year":"2013","journal-title":"Antioxid. Redox Signal."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1212\/WNL.0000000000001718","article-title":"Effect of comorbidity on mortality in multiple sclerosis","volume":"85","author":"Marrie","year":"2015","journal-title":"Neurology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.pharmthera.2017.11.001","article-title":"The sphingosine 1-phosphate receptor modulator fingolimod as a therapeutic agent: Recent findings and new perspectives","volume":"185","author":"Huwiler","year":"2018","journal-title":"Pharmacol. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"104170","DOI":"10.1016\/j.phrs.2019.02.009","article-title":"Targeting the S1P receptor signaling pathways as a promising approach for treatment of autoimmune and inflammatory diseases","volume":"154","author":"Stepanovska","year":"2020","journal-title":"Pharmacol. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3153","DOI":"10.2147\/DDDT.S122249","article-title":"Spotlight on siponimod and its potential in the treatment of secondary progressive multiple sclerosis: The evidence to date","volume":"11","author":"Gajofatto","year":"2017","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1124\/pr.107.07113","article-title":"\u201cInside-out\u201d signaling of sphingosine-1-phosphate: Therapeutic targets","volume":"60","author":"Takabe","year":"2008","journal-title":"Pharmacol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1140","DOI":"10.1111\/j.1476-5381.2011.01260.x","article-title":"Pharmacological relevance and potential of sphingosine 1-phosphate in the vascular system","volume":"163","author":"Schuchardt","year":"2011","journal-title":"Br. J. Pharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1080\/17460441.2019.1646244","article-title":"Preclinical discovery and development of fingolimod for the treatment of multiple sclerosis","volume":"14","author":"Volpi","year":"2019","journal-title":"Expert Opin. Drug Discov."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1038\/ni1545","article-title":"Finding a way out: Lymphocyte egress from lymphoid organs","volume":"8","author":"Schwab","year":"2007","journal-title":"Nat. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1111\/tra.12343","article-title":"Endocytosis of Ligand-Activated Sphingosine 1-Phosphate Receptor 1 Mediated by the Clathrin-Pathway","volume":"17","author":"Reeves","year":"2016","journal-title":"Traffic"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4797","DOI":"10.1111\/bph.12620","article-title":"Investigating the molecular mechanisms through which FTY 720-P causes persistent S1P1 receptor internalization","volume":"171","author":"Sykes","year":"2014","journal-title":"Br. J. Pharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1038\/nrd3248","article-title":"Fingolimod (FTY720): Discovery and development of an oral drug to treat multiple sclerosis","volume":"9","author":"Brinkmann","year":"2010","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_13","first-page":"119","article-title":"Cardiovascular effects of fingolimod: A review article","volume":"13","author":"Behjati","year":"2014","journal-title":"Iran. J. Neurol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1038\/sj.bjp.0706452","article-title":"The immunomodulator FTY720 and its phosphorylated derivative activate the Smad signalling cascade and upregulate connective tissue growth factor and collagen type IV expression in renal mesangial cells","volume":"147","author":"Xin","year":"2006","journal-title":"Br. J. Pharmacol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1016\/j.neuropharm.2014.05.012","article-title":"Novel oxazolo-oxazole derivatives of FTY720 reduce endothelial cell permeability, immune cell chemotaxis and symptoms of experimental autoimmune encephalomyelitis in mice","volume":"85","author":"Imeri","year":"2014","journal-title":"Neuropharmacology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1142","DOI":"10.1016\/j.chembiol.2012.07.016","article-title":"A potent and selective S1P1 antagonist with efficacy in experimental autoimmune encephalomyelitis","volume":"19","author":"Quancard","year":"2012","journal-title":"Chem. Biol."},{"key":"ref_17","first-page":"213","article-title":"A mechanistically novel, first oral therapy for multiple sclerosis: The development of fingolimod (FTY720, Gilenya)","volume":"12","author":"Chun","year":"2011","journal-title":"Discov. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1126\/science.1215904","article-title":"Crystal Structure of a Lipid G Protein-Coupled Receptor","volume":"335","author":"Hanson","year":"2012","journal-title":"Science"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1038\/nature10361","article-title":"Crystal structure of the beta(2) adrenergic receptor-Gs protein complex","volume":"477","author":"Rasmussen","year":"2011","journal-title":"Nature"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1038\/nature13430","article-title":"Visualization of arrestin recruitment by a G-protein-coupled receptor","volume":"512","author":"Shukla","year":"2014","journal-title":"Nature"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1084\/jem.169.2.431","article-title":"Spontaneous recovery of rats from experimental allergic encephalomyelitis is dependent on regulation of the immune system by endogenous adrenal corticosteroids","volume":"169","author":"Macphee","year":"1989","journal-title":"J. Exp. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"9833","DOI":"10.1074\/jbc.M412356200","article-title":"Sphingosine 1-phosphate analogs as receptor antagonists","volume":"280","author":"Davis","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Willis, M.A., Cohen, A., and Cohen, J. (2013). Fingolimod therapy for multiple sclerosis. Seminars in Neurology, Thieme Medical Publishers.","DOI":"10.1055\/s-0033-1343794"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1159\/000453392","article-title":"Acute Anterior Uveitis in a Patient Taking Fingolimod (FTY720) for Multiple Sclerosis","volume":"7","author":"Mack","year":"2016","journal-title":"Case Rep. Ophthalmol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"27351","DOI":"10.1074\/jbc.274.39.27351","article-title":"Differential coupling of the sphingosine 1-phosphate receptors Edg-1, Edg-3, and H218\/Edg-5 to the Gi, Gq, and G12 families of heterotrimeric G proteins","volume":"274","author":"Windh","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.pharmthera.2007.04.006","article-title":"Sphingosine 1-phosphate receptors in health and disease: Mechanistic insights from gene deletion studies and reverse pharmacology","volume":"115","author":"Brinkmann","year":"2007","journal-title":"Pharmacol. Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1016\/j.chembiol.2005.04.019","article-title":"S1P1-selective in vivo-active agonists from high-throughput screening: Off-the-shelf chemical probes of receptor interactions, signaling, and fate","volume":"12","author":"Jo","year":"2005","journal-title":"Chem. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1124\/mol.117.109140","article-title":"S1P1 modulator-induced G\u03b1i Signaling and \u03b2-Arrestin Recruitment Are Both Necessary to Induce Rapid and Efficient Reduction of Blood Lymphocyte Count in vivo","volume":"93","author":"Bolli","year":"2018","journal-title":"Mol. Pharmacol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"206","DOI":"10.4049\/jimmunol.1101537","article-title":"Amelioration of collagen-induced arthritis by a novel S1P1 antagonist with immunomodulatory activities","volume":"188","author":"Fujii","year":"2012","journal-title":"J. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1452","DOI":"10.1089\/neu.2017.5391","article-title":"Sphingosine 1-phosphate receptor subtype 1 as a therapeutic target for brain trauma","volume":"35","author":"Cuzzocrea","year":"2018","journal-title":"J. Neurotrauma"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"9722","DOI":"10.1021\/jm3009508","article-title":"An Oral Sphingosine 1-Phosphate Receptor 1 (S1P1) Antagonist Prodrug with Efficacy in Vivo: Discovery, Synthesis, and Evaluation","volume":"55","author":"Angst","year":"2012","journal-title":"J. Med. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1038\/nchembio804","article-title":"Enhancement of capillary leakage and restoration of lymphocyte egress by a chiral S1P 1 antagonist in vivo","volume":"2","author":"Sanna","year":"2006","journal-title":"Nat. Chem. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Bigaud, M., Dincer, Z., Bollbuck, B., Dawson, J., Beckmann, N., Beerli, C., Fishli-Cavelti, G., Nahler, M., Angst, D., and Janser, P. (2016). Pathophysiological consequences of a break in S1P1-Dependent homeostasis of vascular permeability revealed by S1P1 competitive antagonism. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0168252"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"13","DOI":"10.2147\/CE.S10101","article-title":"Evidence-based assessment of potential use of fingolimod in treatment of relapsing multiple sclerosis","volume":"6","author":"Portaccio","year":"2011","journal-title":"Core Evid."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1021\/cb5008426","article-title":"Drugging sphingosine kinases","volume":"10","author":"Santos","year":"2015","journal-title":"ACS Chem. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1007\/s10620-007-0133-6","article-title":"Suppression of ulcerative colitis in mice by orally available inhibitors of sphingosine kinase","volume":"53","author":"Maines","year":"2008","journal-title":"Dig. Dis. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.antiviral.2018.08.010","article-title":"Transient inhibition of sphingosine kinases confers protection to influenza A virus infected mice","volume":"158","author":"Xia","year":"2018","journal-title":"Antivir. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12864-019-6367-9","article-title":"Genetic deletion of Sphk2 confers protection against Pseudomonas aeruginosa mediated differential expression of genes related to virulent infection and inflammation in mouse lung","volume":"20","author":"Ebenezer","year":"2019","journal-title":"BMC Genom."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Ghosh, M., Thangada, S., Dasgupta, O., Khanna, K.M., Yamase, H.T., Kashgarian, M., Hla, T., Shapiro, L.H., and Ferrer, F.A. (2018). Cell-intrinsic sphingosine kinase 2 promotes macrophage polarization and renal inflammation in response to unilateral ureteral obstruction. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0194053"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1111\/bph.13670","article-title":"Effect of sphingosine kinase modulators on interleukin-1\u03b2 release, sphingosine 1-phosphate receptor 1 expression and experimental autoimmune encephalomyelitis","volume":"174","author":"Barbour","year":"2017","journal-title":"Br. J. Pharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Stepanovska, B., Lange, A.I., Schwalm, S., Pfeilschifter, J.M., Coldewey, S.M., and Huwiler, A. (2020). Downregulation of S1P Lyase Improves Barrier Function in Human Cerebral Microvascular Endothelial Cells Following an Inflammatory Challenge. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21041240"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/0014-5793(94)01132-X","article-title":"Transforming growth factor \u03b22 stimulates acute and chronic activation of the mitogen-activated protein kinase cascade in rat renal mesangial cells","volume":"354","author":"Huwiler","year":"1994","journal-title":"FEBS Lett."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Blanchard, O., Stepanovska, B., Starck, M., Erhardt, M., R\u00f6mer, I., Zu Heringdorf, D.M., Pfeilschifter, J.M., Zangemeister-Wittke, U., and Huwiler, A. (2018). Downregulation of the S1P Transporter Spinster Homology Protein 2 (Spns2) Exerts an Anti-Fibrotic and Anti-Inflammatory Effect in Human Renal Proximal Tubular Epithelial Cells. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19051498"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.neuropharm.2016.01.031","article-title":"Sphingosine kinase 2 deficient mice exhibit reduced experimental autoimmune encephalomyelitis: Resistance to FTY720 but not ST-968 treatments","volume":"105","author":"Imeri","year":"2016","journal-title":"Neuropharmacology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/18\/6463\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:06:46Z","timestamp":1760177206000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/18\/6463"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,4]]},"references-count":44,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["ijms21186463"],"URL":"https:\/\/doi.org\/10.3390\/ijms21186463","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,4]]}}}