S1P1 Receptor Modulation with Cyclical Recovery from Lymphopenia Ameliorates Mouse Model of Multiple Sclerosis

   1. Pedro J. Gonzalez-Cabrera,
   2. Stuart M. Cahalan,
   3. Nhan Nguyen,
   4. Gor Sarkisyan,
   5. Nora B. Leaf,
   6. Michael D. Cameron,
   7. Tomoyuki Kago1 and
   8. Hugh Rosen

   1.
      Departments of Chemical Physiology (P.J.G.C., S.M.C., N.N., G.S., N.B.L., T.K., H.R.) and Immunology (S.M.C.), The Scripps Research Institute, La Jolla, California; and Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, Florida (M.D.C.)

   1. Address correspondence to:
      Dr Hugh Rosen, Department of Chemical Physiology, Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. E-mail: hrosen{at}scripps.edu

   1.

      P.J.G.C. and S.M.C. contributed equally to this work.

    *

      ↵1 Current affiliation: Kyorin Pharmaceutical Co., Pharmacology Division, Tochigi, Japan.

Abstract

Multiple sclerosis (MS) therapies modulate T-cell autoimmunity in the central nervous system (CNS) but may exacerbate latent infections. Fingolimod, a nonselective sphingosine-1-phosphate (S1P) receptor agonist that induces sustained lymphopenia and accumulates in the CNS, represents a new treatment modality for MS. We hypothesized that sustained lymphopenia would not be required for efficacy and that a selective, CNS-penetrant, peripherally short-acting, S1P1 agonist would show full efficacy in a mouse MS model. Using daily treatment with 10 mg/kg 2-(4-(5-(3,4-diethoxyphenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl amino)ethanol (CYM-5442) at the onset of clinical signs in myelin oligodendrocyte glycoprotein MOG35–55- induced experimental allergic encephalomyelitis (EAE), we assessed clinical scores, CNS cellular infiltration, demyelination, and gliosis for 12 days with CYM-5442, vehicle, or fingolimod. CYM-5442 levels in CNS and plasma were determined at experiment termination, and blood lymphopenia was measured 3 and 24 h after the last injection. Plasma levels of cytokines were assayed at the end of the protocol. Changes in S1P1-enhanced green fluorescent protein expression on neurons and astrocytes during active EAE and upon CYM-5442 treatment were quantified with flow cytometry and Western blotting by using native-locus enhanced green fluorescent protein-tagged S1P1 mice. S1P1 agonism alone reduced pathological features as did fingolimod (maximally lymphopenic throughout), despite full reversal of lymphopenia within each dosing interval. CYM-5442 levels in CNS but not in plasma were sustained. Neuronal and astrocytic S1P1 expression in EAE was suppressed by CYM-5442 treatment, relative to vehicle, and levels of key cytokines, such as interleukin 17A, were also significantly reduced in drug-treated mice. S1P1-selective agonists that induce reversible lymphopenia while persisting in the CNS may be effective MS treatments.
Footnotes

    *

      H.R. is a cofounder of Receptos, Inc.
    *

      ↵Graphic The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.
    *

      This work was supported by the National Institutes of Health National Institute of Allergy and Infectious Diseases [Grants U01-AI074564, R01-AI055509] and the National Institutes of Health National Institute of Mental Health [Grant U54-MH084512].
    *

      Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.

      http://dx.doi.org/10.1124/mol.111.076109.
    *

      ABBREVIATIONS:

      MS
          multiple sclerosis
      CNS
          central nervous system
      PBS
          phosphate-buffered saline
      PE
          phycoerythrin
      APC
          antigen-presenting cell
      S1P
          sphingosine-1-phosphate
      MOG
          myelin oligodendrocyte glycoprotein
      EAE
          experimental allergic encephalomyelitis
      eGFP
          enhanced green fluorescent protein
      PTX
          pertussis toxin
      LFB
          Luxol fast blue
      CYM-5442
          2-(4-(5-(3,4-diethoxyphenyl)-1,2,4-oxadiazol-3-yl)-2,3-dihydro-1H-inden-1-yl amino)ethanol
      RP-001
          N-[4-[5-[3-cyano-4-(1-methylethoxy)phenyl]-1,2,4-oxadiazol-3-yl]-2,3-dihydro-1H-iden-1-yl]-β-alanine. 

    * Received September 27, 2011.
    * Accepted October 26, 2011.

    * Copyright © 2012 The American Society for Pharmacology and Experimental Therapeutics

