The immunosuppressant FTY720 down‐regulates sphingosine 1‐phosphate G protein‐coupled receptors

Gräler, Markus H.; Goetzl, Edward J.

doi:10.1096/fj.03-0910fje

Cited by 508 publications

(441 citation statements)

References 41 publications

Supporting

Mentioning

416

Contrasting

Unclassified

Order By: Relevance

“…[1] The FTY720-P-mediated activation of the S1P1 receptor on lymphocytes induces receptor internalization, which attenuates T-cell response to S1P gradients, preventing their egress from secondary lymphoid tissues. [3] In addition to playing a role in the immune system, all S1P receptors except S1P4 are also found differentially expressed in the central nervous system [4] and on various tumor cell types. [5,6] Although the precise regulation of these receptors by locally released S1P remains unclear, S1P receptors are thought to play a role in such events as astrocyte migration, [7] oligodendrocyte differentiation, and cell survival [8,9] and neurogenesis.…”

mentioning

confidence: 99%

Design and Synthesis of Selective and Potent Orally Active S1P5 Agonists

et al. 2010

View full text Add to dashboard Cite

The immunomodulatory drug fingolimod (FTY720, 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol), derived from a fungal metabolite myriocin), is phosphorylated in vivo by sphingosine kinases to produce (R)-FTY720-phosphate (FTY720-P). [1,2] FTY720-P activates sphingosine-1-phosphate (S1P) receptors S1P1, S1P3, S1P4, and S1P5 at low nanomolar concentrations and is inactive toward the S1P2 receptor.[1] The FTY720-P-mediated activation of the S1P1 receptor on lymphocytes induces receptor internalization, which attenuates T-cell response to S1P gradients, preventing their egress from secondary lymphoid tissues. [3] In addition to playing a role in the immune system, all S1P receptors except S1P4 are also found differentially expressed in the central nervous system [4] and on various tumor cell types. [5,6] Although the precise regulation of these receptors by locally released S1P remains unclear, S1P receptors are thought to play a role in such events as astrocyte migration, [7] oligodendrocyte differentiation, and cell survival [8,9] and neurogenesis. [10,11] To assess the relevance of individual S1P receptor subtypes for the activity of FTY720-P, selective agonists are required. Because S1P5 receptors are expressed on oligodendrocytes, and S1P5 receptors are thought to play a role in oligodendrocyte differentiation and survival, we focused on the development of S1P5 agonists. By using a highthroughput screening calcium mobilization assay with GPCR priming and FLIPR technology, [12] we discovered benzamide 1, which has good in vitro potency toward the S1P5 receptor (EC 50 = 270 nm), but has modest selectivity against S1P1 (EC 50 = 3140 nm) and S1P4 (EC 50 = 100 nm). Herein we report our studies of various benzamide modifications carried out to improve the selectivity, bioactivity, pharmacokinetic properties, and ancillary profile of 1, ultimately resulting in the discovery of potent and very selective S1P5 agonists.To guide the optimization process, homology models of all S1P receptors were built from a crystal structure of bovine rhodopsin (PDB ID: 1F88). [13] Docking experiments of 1 into these models revealed a possible location of the binding site, some essential features of the interactions, and indicated potential regions for gaining selectivity and improving potency. In these complexes (Figure 1), 1 adopts a twisted conformation with the aniline ring,~708 out of the benzamide plane and stabilized by a hydrogen bond between the aniline NH group and the amide carbonyl. In the S1P5 receptor complex, the amide group forms a hydrogen bond with OG1-Thr120. The benzamide phenyl ring lies in a large hydrophobic pocket surrounded by Phe196, Phe201, Phe268, Leu119, Trp264, Leu267, and Leu271. The aniline ring undergoes a T-shaped interaction with Phe116 and hydrophobic contacts with Leu271 and Leu292. The ortho-methyl substituents fill a small pocket formed by Tyr89, Val115, and Leu292 on one side, and sit at the face of Phe196 on the other side. Inspection of sequence alignments (Figure 2) revealed two positions, o...

show abstract

mentioning

confidence: 99%

Design and Synthesis of Selective and Potent Orally Active S1P5 Agonists

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Recently, selective inactivation of several S1PR, with the most prominent effect on S1P 1 , has been reported for single S1PR-null cell transductants upon treatment with FTY720. These effects were shown to differ from the classic agonistic functions of the phosphorylated form, but have also added further complexity to the situation [12]. One theory proposes a loss of responsiveness towards S1P in the plasma by FTY720, resulting in inhibition of lymphocyte re-entrance into the circulation.…”

Section: Discussionmentioning

confidence: 99%

“…The current concept of FTY720-induced lymphopenia is that FTY720 modulates lymphocyte trafficking in a dual manner: acceleration of migration into secondary lymphoid organs and blocking the egress into efferent lymphatics. Recent evidence suggests that these differing effects result from selective S1PR inactivation via receptor down-regulation, thereby trapping lymphocytes in secondary lymphatic organs [11,12]. In vascular endothelial cells, FTY720-P antagonizes VEGF-induced vascular permeability by inducing adherens junction assembly [13].…”

Section: Introductionmentioning

confidence: 99%

The immunomodulator FTY720 interferes with effector functions of human monocyte‐derived dendritic cells

et al. 2005

View full text Add to dashboard Cite

The potent immunomodulator FTY720 elicits immunosuppression via acting on sphingosine 1-phosphate receptors (S1PR), thereby leading to an entrapment of lymphocytes in the secondary lymphoid tissue. To elucidate the potential in vitro effects of this drug on human monocyte-derived DC, we used low nanomolar therapeutic concentrations of FTY720 and phosphorylated FTY720 (FTY720-P) and investigated their influence on DC surface marker expression, protein levels of S1PR and DC effector functions: antigen uptake, chemotaxis, cytokine production, allostimulatory and Thpriming capacity. We report that both FTY720 and FTY720-P reduce chemotaxis of immature and mature DC. Mature DC generated in the presence of FTY720 or FTY720-P showed an impaired immunostimmulatory capacity and reduced IL-12 but increased IL-10 production. T cells cultured in the presence of FTY720-or FTY720-P-treated DC showed an altered cytokine production profile indicating a shift from Th1 toward Th2 differentiation. In treated immature and mature DC, expression levels for two S1PR proteins, S1P 1 and S1P 4, were reduced. We conclude that in vitro treatment with FTY720 affects DC features that are essential for serving their role as antigen-presenting cells. This might represent a new aspect of the overall immunosuppressive action of FTY720 and makes DC potential targets of further sphingolipid-derived drugs.

show abstract

“…Currently, the discussion about its mode of action is mainly focused on its property to cause lymph node homing or sequestration of T cells. In this scenario, FTY720 activates sphingosine-1-phosphate (S1P) 4 receptors and modulates migration after being effectively phosphorylated in vivo by sphingosine kinase 2 (SphK2) (5,6). FTY720-phosphate (FTY720-P) exhibits a potency comparable to S1P itself as an agonist at four of the five known G-protein-coupled S1PRs (S1P 1,3,4,5 ).…”

mentioning

confidence: 99%

“…In this scenario, FTY720 activates sphingosine-1-phosphate (S1P) 4 receptors and modulates migration after being effectively phosphorylated in vivo by sphingosine kinase 2 (SphK2) (5,6). FTY720-phosphate (FTY720-P) exhibits a potency comparable to S1P itself as an agonist at four of the five known G-protein-coupled S1PRs (S1P 1,3,4,5 ). Interference of FTY720 with S1P signaling hampers entry of lymphocytes into efferent lymphatics within lymph nodes, thereby delaying their subsequent return into circulation (7,8).…”

mentioning

confidence: 99%

FTY720 Ameliorates Th1-Mediated Colitis in Mice by Directly Affecting the Functional Activity of CD4+CD25+ Regulatory T Cells

Daniel

Sartory

Zahn

et al. 2007

The Journal of Immunology

155

122

View full text Add to dashboard Cite

Following the present concepts, the synthetic sphingosine analog of myriocin FTY720 alters migration and homing of lymphocytes via sphingosine 1-phosphate receptors. However, several studies indicate that the immunosuppressive properties of FTY720 may alternatively be due to tolerogenic activities via modulation of dendritic cell differentiation or based on direct effects on CD4+CD25+ regulatory T cells (Treg). As Treg play an important role for the cure of inflammatory colitis, we used the Th1-mediated 2,4,6-trinitrobenzene sulfonic acid (TNBS) colitis model to address the therapeutic potential of FTY720 in vivo. A rectal enema of TNBS was given to BALB/c mice. FTY720 was administered i.p. from days 0 to 3 or 3 to 5. FTY720 substantially reduced all clinical, histopathologic, macroscopic, and microscopic parameters of colitis analyzed. The therapeutic effects of FTY720 were associated with a down-regulation of IL-12p70 and subsequent Th1 cytokines. Importantly, FTY720 treatment resulted in a prominent up-regulation of FoxP3, IL-10, TGFβ, and CTLA4. Supporting the hypothesis that FTY720 directly affects functional activity of CD4+CD25+ Treg, we measured a significant increase of CD25 and FoxP3 expression in isolated lamina propria CD4+ T cells of FTY720-treated mice. The impact of FTY720 on Treg induction was further confirmed by concomitant in vivo blockade of CTLA4 or IL-10R which significantly abrogated its therapeutic activity. In conclusion, our data provide clear evidence that in addition to its well-established effects on migration FTY720 leads to a specific down-regulation of proinflammatory signals while simultaneously inducing functional activity of CD4+CD25+ Treg. Thus, FTY720 may offer a promising new therapeutic strategy for the treatment of IBD.

show abstract

The immunosuppressant FTY720 down‐regulates sphingosine 1‐phosphate G protein‐coupled receptors

Cited by 508 publications

References 41 publications

Design and Synthesis of Selective and Potent Orally Active S1P5 Agonists

Design and Synthesis of Selective and Potent Orally Active S1P5 Agonists

The immunomodulator FTY720 interferes with effector functions of human monocyte‐derived dendritic cells

FTY720 Ameliorates Th1-Mediated Colitis in Mice by Directly Affecting the Functional Activity of CD4+CD25+ Regulatory T Cells

Contact Info

Product

Resources

About