Selective Sphingosine 1-phosphate Research Articles

Siponimod for multiple sclerosis

Siponimod for multiple sclerosis

Central Modulation of Selective Sphingosine-1-Phosphate Receptor 1 Ameliorates Experimental Multiple Sclerosis

Future treatments of multiple sclerosis (MS), a chronic autoimmune neurodegenerative disease of the central nervous system (CNS), aim for simultaneous early targeting of peripheral immune function and neuroinflammation. Sphingosine-1-phosphate (S1P) receptor modulators are among the most promising drugs with both “immunological” and “non-immunological” actions. Selective S1P receptor modulators have been recently approved for MS and shown clinical efficacy in its mouse model, the experimental autoimmune encephalomyelitis (EAE). Here, we investigated the anti-inflammatory/neuroprotective effects of ozanimod (RPC1063), a S1P1/5 modulator recently approved in the United States for the treatment of MS, by performing ex vivo studies in EAE brain. Electrophysiological experiments, supported by molecular and immunofluorescence analysis, revealed that ozanimod was able to dampen the EAE glutamatergic synaptic alterations, through attenuation of local inflammatory response driven by activated microglia and infiltrating T cells, the main CNS-cellular players of EAE synaptopathy. Electrophysiological studies with selective S1P1 (AUY954) and S1P5 (A971432) agonists suggested that S1P1 modulation is the main driver of the anti-excitotoxic activity mediated by ozanimod. Accordingly, in vivo intra-cerebroventricular treatment of EAE mice with AUY954 ameliorated clinical disability. Altogether these results strengthened the relevance of S1P1 agonists as immunomodulatory and neuroprotective drugs for MS therapy.

Etrasimod, a Selective S1P1,4,5 Receptor Modulator, Exhibits Differential G Protein Signaling Compared to Other S1P Receptor Modulators

BackgroundEtrasimod (APD334) is an oral, selective sphingosine‐1‐phosphate (S1P) receptor modulator in development for atopic dermatitis and inflammatory bowel disease. In humans, S1P receptor modulators decrease heart rate at first‐dose due to initial on‐target S1P receptor 1 (S1P1) agonism in cardiac tissue. S1P1 agonism activates Gi and β‐arrestin signaling pathways: β‐arrestin activation leads to receptor internalization, the proposed mechanism by which S1P receptor modulators regulate lymphocyte trafficking; Gi‐coupled signaling activates G‐protein coupled inwardly rectifying potassium (GIRK) channels, that may contribute to the regulation of heart rate. Approved or investigational S1P receptor modulators have demonstrated different degrees of heart rate reduction and atrioventricular block in clinical studies. Such differences may be the result of differential signaling through the Gi coupled GIRK pathway.AimTo investigate S1P receptor expression in human sinoatrial (SA) node tissue, and the relative S1P1 mediated signaling events and GIRK channel activation of etrasimod and other S1P receptor modulators in the context of first dose heart rate effects.MethodsS1P receptor profiling was performed by qPCR in normal human primary SA node and right atrium samples. Potencies for multiple S1P receptor modulators to activate different S1P1 downstream signaling pathways (GTPγS, cAMP, β‐arrestin, S1P1 internalization) were measured in recombinant human‐S1P1 expressing cells. Gi mediated inhibition of cAMP was also measured in primary HUVEC. GIRK channel activation was measured in isolated adult human primary atrial myocytes.ResultsProfiling of S1P receptors in normal human atrial tissue and SA node demonstrated enriched expression of S1P1,2,3 with only minor expression of S1P4,5. Etrasimod was equipotent to other S1P receptor‐ modulators such as ozanimod, fingolimod, and siponimod in promoting β‐arrestin recruitment and S1P1 receptor internalization. Etrasimod was significantly less potent than ozanimod, fingolimod, and siponimod in assays measuring G‐protein activation (GTPγS binding and cAMP inhibition). In comparisons of GIRK activation, etrasimod was found to be a less potent activator of human atrial GIRK channels than ozanimod.ConclusionsWhen compared to other S1P1 modulators, the pharmacological profile of etrasimod at the S1P1 receptor suggests diminished capacity for signaling through Gi‐protein dependent pathways believed to be responsible for transient reductions in heart rate, while maintaining potent signaling in the β‐arrestin recruitment and receptor internalization related pathways underpinning the therapeutic efficacy of S1P1 modulators in immune‐mediated inflammatory disease settings.Support or Funding InformationThe study was sponsored by Arena Pharmaceuticals, Inc.

Amiselimod (MT-1303), a Novel Sphingosine 1-Phosphate Receptor-1 Modulator, Potently Inhibits the Progression of Lupus Nephritis in Two Murine SLE Models.

Amiselimod (MT-1303) is a novel and selective sphingosine 1-phosphate receptor-1 (S1P1) modulator with a more favorable cardiac safety profile than other S1P1 receptor modulators. In this study, we evaluated the effects of MT-1303 on the progression of lupus nephritis in two well-known murine systemic lupus erythematosus (SLE) models, MRL/lpr and NZBWF1 mice, compared with those of FK506. Daily oral doses of 0.1 and 0.3 mg/kg MT-1303 not only inhibited the development of lupus nephritis when administered before onset in MRL/lpr and NZBWF1 mice but also improved symptoms of lupus nephritis when administered after onset in MRL/lpr mice. Its efficacy in these models was more potent or comparable to that of FK506 (1 and 3 mg/kg). In histological analysis, treatment with MT-1303 inhibited infiltration of T cells into the kidneys, mesangial expansion, and glomerular sclerosis. MT-1303 treatment resulted in a marked reduction in T cells and B cells in the peripheral blood and significantly inhibited increases in the number of plasma cells in the spleen and T cells in the kidneys. In addition, administration of MT-1303 suppressed elevations in serum anti-dsDNA antibody levels in MRL/lpr mice, but not in NZBWF1 mice. Our findings show that MT-1303 exhibits marked therapeutic effects on lupus nephritis in two SLE models, likely by reducing the infiltration of autoreactive T cells into the kidneys. These results suggest that MT-1303 has the potential to be used as a therapeutic agent for patients suffering from SLE, including lupus nephritis.

Efficacy and Safety of Etrasimod in a Phase 2 Randomized Trial of Patients With Ulcerative Colitis

Etrasimod (APD334) is an oral, selective sphingosine 1-phosphate receptor modulator in development for immune-mediated inflammatory disorders. We assessed the efficacy and safety of etrasimod in patients with moderately to severely active ulcerative colitis (UC). In a phase 2, proof-of-concept, double-blind, parallel-group study, adult outpatients with modified Mayo Clinic scores (MCSs) (stool frequency, rectal bleeding, and endoscopy findings) of 4-9, endoscopic subscores of 2 or more, and rectal bleeding subscores of 1 or more were randomly assigned to groups given once-daily etrasimod 1 mg (n= 52), etrasimod 2 mg (n= 50), or placebo (n= 54) for 12 weeks. The study was performed from October 15, 2015, through February 14, 2018, at 87 centers in 17 countries. The primary endpoint was an increase in the mean improvement in modified MCS from baseline to week 12. Secondary endpoints included the proportion of patients with endoscopic improvement (subscores of 1 or less) from baseline to week 12. Exploratory endpoints, including clinical remission, are reported in the article, although the study was statistically powered to draw conclusions only on the primary endpoint. At week 12, the etrasimod 2 mg group met the primary and all secondary endpoints. Etrasimod 2 mg led to a significantly greater increase in mean improvement in modified MCS from baseline than placebo (difference from placebo, 0.99 points; 90% confidence interval, 0.30-1.68; P= .009), and etrasimod 1 mg led to an increase in mean improvement from baseline in modified MCS of 0.43 points more than placebo (90% confidence interval, reduction of 0.24 to increase of 1.11; nominal P= .15). Endoscopic improvement occurred in 41.8% of patients receiving etrasimod 2 mg vs 17.8% receiving placebo (P= .003). Most adverse events were mild to moderate. Three patients had a transient, asymptomatic, low-grade atrioventricular block that resolved spontaneously all patients had evidence of atrioventricular block before etrasimod exposure. In patients with moderately to severely active ulcerative colitis, etrasimod 2 mg was more effective than placebo in producing clinical and endoscopic improvements. Further clinical development is warranted. Clinicaltrials.gov, Number: NCT02447302.

First use of cenerimod, a selective S1P1 receptor modulator, for the treatment of SLE: a double-blind, randomised, placebo-controlled, proof-of-concept study

ObjectiveTo investigate the pharmacodynamics, pharmacokinetics and safety of cenerimod—a potent, oral, selective sphingosine 1-phosphate 1 receptor modulator—in patients with SLE.MethodsThis multicentre, double-blind, placebo-controlled study was conducted in two parts. In...

030 Etrasimod, an oral, selective sphingosine 1-phosphate receptor modulator improves skin inflammation in a contact hypersensitivity dermatitis model

Etrasimod (APD334) is a selective sphingosine 1-phosphate receptor 1,4,5 (S1P1,4,5) modulator in development for atopic dermatitis (AD). S1P1 modulates trafficking of many immune cells, including those in eczematous skin. This study evaluated the effect of etrasimod on circulating and tissue immune cells and its impact on skin inflammation in a dermatitis mouse model. BALB/c mice were epicutaneously sensitized with fluorescein isothiocyanate (FITC) on the hind flank on Days (D) 0 and 5, then epicutaneously FITC challenged on the ear on D10-12. From D -1, mice were dosed orally once daily with vehicle, dexamethasone (Dex), or etrasimod (1 or 3 mg/kg). Ear thickness was measured with calipers on D0, 5, 10-13. On D2 and 13, draining lymph nodes (dLN) were analyzed by flow cytometry (FACS) for cellularity and activation. On D13, we examined blood for immune cell count and cytokines, and ear skin by FACS and histology. Sensitization did not cause skin inflammation, yet on D2 dendritic cells (DC), T cells and B cells expanded in the dLN in all mice. In vehicle mice, challenges amplified skin thickening from D10-13, and terminal analyses revealed worse histologic score and immune cell expansion in dLN and ear skin. Etrasimod dose-dependently lessened ear thickening and histologic score, with the high dose achieving comparable efficacy as Dex on D13. Etrasimod reduced lymphocyte frequency in the blood. In the dLN, etrasimod reduced DC influx, and decreased expansion and activation of T cells. B cells and eosinophils were also reduced in number. In the ear skin, the increase of T cells, B cells, and eosinophils was dose-dependently inhibited. In conclusion, etrasimod effectively reduced ear skin inflammation and dermatitis in FITC-induced hypersensitivity. Etrasimod significantly reduced the activation and expansion of immune cells after challenge in the dLN and ear skin. Notably, the dose-dependent reduction of immune cells in ear skin correlated with improvements in disease. This data encourages further study of etrasimod as a novel therapy for AD.

The selective sphingosine 1-phosphate receptor 1 modulator RP101075 improves microvascular circulation after cerebrovascular thrombosis.

Sphingosine-1-phosphate receptor (S1PR) modulators provide protection in preclinical and clinical studies for ischemic stroke, but the influences of S1PR modulation on microvascular thrombosis remain poorly understood. This study investigates the impact of a selective S1PR1 modulator RP101075 on microvascular circulation in a mouse model of laser-induced thrombosis. The flow velocity of cortical arterioles in mice was measured in vivo under 2-photon laser scanning microscopy. Thrombosis was induced in cortical arterioles by laser irritation. At 30 min after laser-induced thrombosis, mice were treated with either RP101075 or vehicle. RP101075 did not alter the flow velocity of cortical arterioles under physiologic conditions. Laser-induced thrombosis led to a pronounced reduction of flow velocity in cortical arterioles that persisted for ≥90 min. The reduction of flow velocity in cortical arterioles following thrombosis was significantly attenuated following RP101075 treatment. RP101075 did not significantly affect coagulation time, bleeding time, heart rate, and blood pressure. In addition, RP101075 treatment reduced thrombus volume, which was accompanied by a reduction of leukocyte content in the thrombus. Our findings demonstrate that the selective S1PR1 modulator RP101075 improves microvascular circulation after thrombosis, implying a component of improved microvascular circulation to the benefit of S1PR modulation in cerebral ischemia.-Li, H., Zhou, X., Li, Y., Ma, X., Gonzales, R. J., Qiu, S., Shi, F.-D., Liu, Q. The selective sphingosine 1-phosphate receptor 1 modulator RP101075 improves microvascular circulation after cerebrovascular thrombosis.

Siponimod: First Global Approval.

Siponimod (Mayzent®) is an oral selective sphingosine 1-phosphate receptor subtypes 1 and 5 (S1PR1,5) modulator being developed by Novartis Pharmaceuticals for the treatment of multiple sclerosis (MS) and intracerebral haemorrhage. In March 2019, siponimod received its first global approval in the USA, for the treatment of adults with relapsing forms of MS, including clinically isolated syndrome, relapsing-remitting disease and active secondary progressive disease. Siponimod is under regulatory review in the EU and Japan for secondary progressive MS. This article summarizes the milestone in the development of siponimod leading to this first global approval for MS in the USA.

APD588, a novel, selective S1P- receptor modulator, inhibits inflammation and airway hyperresponsiveness in the ovalbumin-induced model of allergic asthma

Abstract Background FTY720 is a non-selective sphingosine-1-phosphate (S1P) receptor modulator that has shown efficacy in mouse asthma models. APD588 is an investigational stage S1P receptor modulator with increased selectivity and potential for an improved safety profile. We hypothesized that the optimized S1P receptor profile of APD588 would confer beneficial effects in ovalbumin (OVA)-induced asthma. Aim To evaluate efficacy of prophylactic APD588 in the OVA-induced allergic asthma model Methods Receptor potency and selectivity profile of APD588 was determined (b-arrestin, GTPgS) in cell lines expressing recombinant human S1P1–5. Efficacy in allergic asthma was assessed in BALB/c mice which were sensitized with intraperitoneal OVA on days 0, 7, and 17, then exposed to intratracheal (IT) vehicle or OVA on days 21–23. From day 20, vehicle, APD588 or FTY720 were dosed IT, QD. On day 24, methacholine-induced airway hyperresponsiveness (AHR), immune cell infiltration and BAL cytokines were measured. Results APD588 was a potent modulator of S1P1, with less potency at S1P4, 5, and no detectable activity at S1P2,3. IT administration of APD588 produced a dose-dependent inhibition of AHR, significantly inhibited airway eosinophil and lymphocyte infiltration, and BAL cytokines IL-4 and IL-5. FTY720 also reduced AHR and neutrophil and lymphocyte BAL cell counts, but had no effect on eosinophil counts or cytokines. Conclusions APD588 was an effective IT therapy for OVA-induced asthma. Lung function improvements correlated with reduced Th2 cytokines and lymphocyte and eosinophil infiltration in the BAL. Based on these results, APD588 may have therapeutic potential in asthma, and broadly indicate potential in other atopic Th2 diseases.

Ozanimod for the treatment of relapsing remitting multiple sclerosis

ABSTRACT Introduction Ozanimod is a selective sphingosine 1-phosphate receptor 1 and 5 modulator under development by Celgene, for the treatment of relapsing remitting multiple sclerosis. Extensive clinical experience has become available for the related compound fingolimod, favoring the sphingosine 1-phosphate therapeutic concept. Off-target effects have been attributed to its low receptor specificity and have prompted the development of next generation sphingosine 1-phosphate receptor modulators. Areas covered The authors evaluate the literature of ozanimod, using the PubMed database as well as repositories of the European Committee for Treatment and Research in Multiple Sclerosis and the American and European Academy of Neurology. Specifically, the authors cover and discuss the preclinical data on ozanimod, pharmacokinetics and dynamics, and data on efficacy and safety from the pivotal trials. Expert opinion Superiority of ozanimod over intramuscular interferon β-1a with regard to reduction in annualized relapse rate and magnetic resonance imaging outcomes has been shown in two phase III trials. The beneficial effect on brain volume and gray matter loss are encouraging and in line with data on other newer immunomodulators. Ozanimod is a valuable contribution to the therapeutic armamentarium in MS, although the effect on disability progression is unclear and requires further investigations.

Sphingosine-1-phosphate promotes the proliferation and attenuates apoptosis of Endothelial progenitor cells via S1PR1/S1PR3/PI3K/Akt pathway.

Sphingosine-1-phosphate (S1P) is a bioactive lysophospholipid that involves in numerous pathophysiological processes. Endothelial progenitor cells (EPCs) play a crucial role in endothelial repair and tumor angiogenesis. The aim of study was to determine the effects of S1P on proliferation and anti-apoptosis of EPCs and their signaling pathways. In this study, we showed that S1P, SEW2871 (a selective S1P receptor 1 (S1PR1) agonist), or CYM5541 (a selective S1P receptor 3 (S1PR3) allosteric agonist promotes the proliferation and attenuates apoptosis of bone marrow (BM)-derived EPCs. Futhermore, it was showed that S1P could promote EPCs proliferation, which could be significantly inhibited by pretreatment with CAY10444 (an S1PR3 antagonist), VPC23019 (a selective S1PR(1)/S1PR(3) antagonist), or LY294002 (a PI3K inhibitor). Moveover, we discovered that S1P could significantly attenuate H2 O2 -induced apoptosis and activation of caspase-3 in vitro, while W146 (an S1PR1 antagonist), VPC23019, or LY294002 could significantly increase the activation of caspase-3 and subsequent augmented apoptosis. Our results indicated that the protective effect of S1P is mediated by activating the PI3K/Akt pathway. In addition, S1P promotion of EPCs proliferation was observed to be mainly mediated through S1PR3 and attenuation of EPCs apoptosis induced by H2 O2 was mainly mediated through S1PR1; both of these effects are mediated by activating the PI3K/Akt pathway, which provides potentially useful therapeutic targets for coronary artery disease, diabetes mellitus, and cancer treatment.

Siponimod versus placebo in secondary progressive multiple sclerosis (EXPAND): a double-blind, randomised, phase 3 study

No treatment has consistently shown efficacy in slowing disability progression in patients with secondary progressive multiple sclerosis (SPMS). We assessed the effect of siponimod, a selective sphingosine 1-phosphate (S1P) receptor1,5 modulator, on disability progression in patients with SPMS. This event-driven and exposure-driven, double-blind, phase 3 trial was done at 292 hospital clinics and specialised multiple sclerosis centres in 31 countries. Using interactive response technology to assign numbers linked to treatment arms, patients (age 18-60 years) with SPMS and an Expanded Disability Status Scale score of 3·0-6·5 were randomly assigned (2:1) to once daily oral siponimod 2 mg or placebo for up to 3 years or until the occurrence of a prespecified number of confirmed disability progression (CDP) events. The primary endpoint was time to 3-month CDP. Efficacy was assessed for the full analysis set (ie, all randomly assigned and treated patients); safety was assessed for the safety set. This trial is registered with ClinicalTrials.gov, number NCT01665144. 1651 patients were randomly assigned between Feb 5, 2013, and June 2, 2015 (1105 to the siponimod group, and 546 to the placebo group). One patient did not sign the consent form, and five patients did not receive study drug, all of whom were in the siponimod group. 1645 patients were included in the analyses (1099 in the siponimod group and 546 in the placebo). At baseline, the mean time since first multiple sclerosis symptoms was 16·8 years (SD 8·3), and the mean time since conversion to SPMS was 3·8 years (SD 3·5); 1055 (64%) patients had not relapsed in the previous 2 years, and 918 (56%) of 1651 needed walking assistance. 903 (82%) patients receiving siponimod and 424 (78%) patients receiving placebo completed the study. 288 (26%) of 1096 patients receiving siponimod and 173 (32%) of 545 patients receiving placebo had 3-month CDP (hazard ratio 0·79, 95% CI 0·65-0·95; relative risk reduction 21%; p=0·013). Adverse events occurred in 975 (89%) of 1099 patients receiving siponimod versus 445 (82%) of 546 patients receiving placebo; serious adverse events were reported for 197 (18%) patients in the siponimod group versus 83 (15%) patients in the placebo group. Lymphopenia, increased liver transaminase concentration, bradycardia and bradyarrhythmia at treatment initiation, macular oedema, hypertension, varicella zoster reactivation, and convulsions occurred more frequently with siponimod than with placebo. Initial dose titration mitigated cardiac first-dose effects. Frequencies of infections, malignancies, and fatalities did not differ between groups. Siponimod reduced the risk of disability progression with a safety profile similar to that of other S1P modulators and is likely to be a useful treatment for SPMS. Novartis Pharma AG.

P573 Safety and immune modulatory properties of etrasimod (APD334), a next-generation oral, selective sphingosine 1-phosphate receptor (S1PR) modulator, in healthy volunteers

P573 Safety and immune modulatory properties of etrasimod (APD334), a next-generation oral, selective sphingosine 1-phosphate receptor (S1PR) modulator, in healthy volunteers

Modulation of sphingosine-1-phosphate in inflammatory bowel disease.

Inflammatory bowel diseases (IBD), including ulcerative colitis and Crohn's disease, involve an inappropriate immune reaction in the digestive tract, causing a variety of disabling symptoms. The advent of monoclonal antibodies (anti-tumor necrosis factor, anti-integrin, anti-interleukin -23) has revolutionized IBD management. Nevertheless, these agents, with potential for immunogenicity, are associated with high rates of response loss and disease relapse over time. They are also associated with high production costs. Sphingosine-1-phosphate (S1P), a membrane-derived lysophospholipid signaling molecule, is implicated in a vast array of physiological and pathophysiological processes, primarily via extracellular activation of S1P1-S1P5 receptors. S1P1, S1P4 and S1P5 are involved in regulation of the immune system, while S1P2 and S1P3 may be associated with cardiovascular, pulmonary, and theoretical cancer-related risks. Targeting S1P receptors for inflammatory conditions has been successful in clinical trials leading to approval of the non-selective S1P modulator, fingolimod, for relapsing forms of multiple sclerosis. However, the association of this non-selective S1P modulator with serious adverse events provides the rationale for developing more selective S1P receptor modulators. Until recently, three S1P modulators with differing selectivity for S1P receptors were in clinical development for IBD: ozanimod (RPC1063), etrasimod (APD334) and amiselimod (MT-1303). The development of amiselimod has been stopped as Biogen are currently focusing on other drugs in its portfolio. Following encouraging results from the Phase 2 TOUCHSTONE trial, a Phase 3 trial of the S1P modulator ozanimod in patients with moderate-to-severe ulcerative colitis is ongoing. Etrasimod is also being tested in a phase 2 trial in ulcerative colitis. These pipeline medications can be administered orally and may avoid the formation of anti-drug antibodies that can lead to treatment failure with injectable biologic therapies for IBD. Data from ongoing clinical trials will establish the relationship between the selectivity of S1P modulators and their safety and efficacy in IBD, as well as their potential place in the clinical armamentarium for IBD.

Evaluation of species difference in peripheral lymphocyte reduction effect of CS-0777, a sphingosine 1-phosphate receptor modulator, based on a pharmacokinetic/pharmacodynamic model analysis.

Pharmacokinetic (PK) and pharmacodynamic (PD) modeling was conducted for the reduction of peripheral lymphocytes after oral administration of CS-0777 to healthy rats, monkeys and experimental autoimmune encephalomyelitis (EAE) induced rats. The phosphorylated active metabolite of CS-0777, M1, is a selective sphingosine 1-phosphate receptor-1 modulator. A linear one- and two-compartment model with a reversible metabolism process characterized the time courses of CS-0777 and M1 concentrations in rats and monkeys, respectively. The relationship between lymphocyte counts and M1 concentrations in blood was well described by an indirect response model in all animals examined. An Imax of 0.815 and an IC50 of 6.58nM in healthy rats, an Imax of 0.807 and an IC50 of 5.09nM in the EAE rats, an Imax of 0.789 and an IC50 of 0.484nM in monkeys were estimated by the indirect PD model. Since the IC50 values calculated in terms of the unbound plasma concentration in rats and monkeys were within a similar range, after correction of the IC50 in blood described above with the blood to plasma concentration ratio and the plasma free fraction of M1, it was revealed that there is no species difference in the essential activity of M1 against lymphocyte reduction. The sensitivity of the lymphocytes to M1 was not affected by the EAE status. Comparison of the simulated lymphocyte reduction in EAE rats after multiple dosing with CS-0777 and the actual EAE clinical scores implies that the significant suppressive effect on EAE did not require the elimination of all lymphocytes from the blood. Copyright © 2016 John Wiley & Sons, Ltd.

Safety and Efficacy of Long-Term Treatment with Ozanimod, an Oral S1P Receptor Modulator, in Moderate to Severe Ulcerative Colitis: Touchstone Extension

Introduction: Ozanimod (RPC1063) is an oral, selective sphingosine 1-phosphate (S1P) 1 and 5 receptor modulator in clinical development for the treatment of Inflammatory Bowel Disease (ulcerative colitis [UC] and Crohn's disease) and relapsing multiple sclerosis. The objective of the open-label extension (OLE) of the TOUCHSTONE trial is to evaluate the long-term efficacy and safety of daily 1 mg ozanimod in patients with moderate to severe UC who had initially participated in the TOUCHSTONE trial. Methods: A total of 197 patients were randomized (1:1:1) and treated with daily ozanimod at 0.5 mg, 1mg (n=67), or placebo in the TOUCHSTONE trial. Of the initial 197 patients randomized, 170 (86%) entered the OLE and received daily 1 mg ozanimod, 131 (77%) completed the Week 44 assessments, and 109 (64%) remained in the OLE as of the data cut-off (March 2016). At the data cut-off, all remaining patients had received treatment in the OLE for ≥ 1 year. Results: At entry into the OLE, the partial Mayo Score (pMS) for patients on placebo, ozanimod 0.5 mg, and 1.0 mg was 4.6, 4.5, and 3.3 respectively. The pMS had improved in all groups (1.7, 1.7, and 1.9) at Week 44. The greatest improvement was reported in patients who received placebo or ozanimod 0.5 mg in the TOUCHSTONE trial with a change in pMS at Week 44 of -2.6, -2.7 and -1.3 in the placebo, 0.5 mg and 1 mg groups. Improvement occurred rapidly, in the first 4 to 8 weeks of the OLE with a change in pMS at Week 4 of -1.8, -1.5 and -0.8 and a change in pMS at Week 8 of -2.4, -1.9 and -1.1 in the placebo, 0.5 mg and 1 mg groups. At the Week 44 visit in the OLE, 119/131 (90.9%) had little or no active disease based on the physician global assessment (PGA 0 or 1), 129/131 (98.4%) had little or no blood in their stools (rectal bleeding subscore [RBS] 0 or 1), 111/131 (84.7%) had no blood in the stools (RBS 0). The most common adverse events (AEs) (>2.0%) during OLE were UC flare, anemia, upper respiratory tract infection, nasal pharyngitis, back pain, arthralgia, headache, alanine aminotransferase (ALT) or aspartate aminotransferase (AST) elevation. The only serious AEs in ≥2 patients were anemia, and ulcerative colitis flare. ALT and AST > 3x upper limit of normal occurred in 4 (2.4%) of the 170 patients in the OLE. All elevations were asymptomatic, < 5xULN, transient, and resolving while receiving continued treatment. Conclusion: Long-term treatment with ozanimod continues to be safe and well tolerated with good compliance and evidence of durable efficacy.

Effects of High- and Low-Fat Meals on the Pharmacokinetics of Ozanimod, a Novel Sphingosine 1-Phosphate Receptor Modulator

Introduction: Ozanimod (RPC1063) is an oral, selective sphingosine 1-phosphate (S1P) 1 and 5 receptor modulator in clinical development for the treatment of relapsing multiple sclerosis and inflammatory bowel disease. Since food has the potential to significantly influence the rate and extent of drug absorption, we designed and conducted a food-effect study with ozanimod. Methods: The objective of this study was to characterize the effects of high-fat and low-fat meals on the pharmacokinetics (PK) of a single oral dose of ozanimod in healthy adult subjects. This was a randomized, open-label, 3-period, 6-sequence, crossover study. Twenty-four healthy adult subjects were enrolled to receive a single 1-mg oral dose of ozanimod under 3 different conditions separated by washout periods of 7 days: fasted (treatment A), with a standard FDA high-fat meal (treatment B), and with a low-fat meal (treatment C). PK parameters for ozanimod and its two active metabolites (RP101988 [major] and RP101075 [minor]) were calculated. Point estimates and 90% confidence intervals (CIs) about the geometric mean ratio between treatments for maximum concentration (Cmax) and overall exposure (AUC0-inf or AUC0-last) were determined using linear mixed-effects models. Results: Twenty subjects completed both treatments A and B and 23 subjects completed both treatments A and C and provided PK data for the pair-wise comparison of fed vs fasted. The 90% CIs between fed (high-fat or low-fat) and fasted treatments for the Cmax and AUC0-inf of ozanimod and RP101988 were all within the no-effect boundary of 0.80 to 1.25. For the minor metabolite RP101075, the 90% CIs between high-fat and fasted for AUC0-last and the 90% CIs between low-fat and fasted for Cmax and AUC0-last were within the no-effect boundary. While the 90% CI between high-fat and fasted treatments for RP101075 Cmax (0.76 to 0.88) fell outside the lower bound of the no-effect boundary, this decrease was not considered to be clinically meaningful. Median time to reach Cmax (Tmax) for ozanimod was delayed following a high-fat meal (12 hours) compared to fasted and lowfat meal conditions (8 hours), but the range (6 to 12 hours) was the same. Mean elimination half-life (t1/2) values for ozanimod and its active metabolites ranged from 17-21 hours and were similar across all 3 treatments. Overall, single oral doses of 1 mg ozanimod were considered generally well tolerated. Conclusion: Ozanimod may be taken with or without food.

Sphingosine-1-Phosphate (S1P) and S1P Signaling Pathway: Therapeutic Targets in Autoimmunity and Inflammation.

Sphingosine-1-phosphate (S1P) and S1P receptors (S1PR) are ubiquitously expressed. S1P-S1PR signaling has been well characterized in immune trafficking and activation in innate and adaptive immune systems. However, the full extent of its involvement in the pathogenesis of autoimmune diseases is not well understood. FTY720 (fingolimod), a non-selective S1PR modulator, significantly decreased annualized relapse rates in relapsing-remitting multiple sclerosis (MS). FTY720, which primarily targets S1P receptor 1 asafunctional antagonist, arrests lymphocyte egress from secondary lymphoid tissues and reduces neuroinflammation in the central nervous system (CNS). Recent studies suggest that FTY720 also decreases astrogliosis and promotes oligodendrocyte differentiation within the CNS and may have therapeutic benefit to prevent brain atrophy. Since S1P signaling is involved in multiple immune functions, therapies targeting S1P axis may be applicable to treat autoimmune diseases other than MS. Currently, over a dozen selective S1PR and S1P pathway modulators with potentially superior therapeutic efficacy and better side-effect profiles are in the pipeline of drug development. Furthermore, newly characterized molecules such as apolipoprotein M (ApoM) (S1P chaperon) and SPNS2 (S1P transporter) are also potential targets for treatment of autoimmune diseases. Finally, the application of therapies targeting S1P and S1P signaling pathways may be expanded to treat several other immune-mediated disorders (such as post-infectious diseases, post-stroke and post-stroke dementia) and inflammatory conditions beyond their application in primary autoimmune diseases.

Safety and efficacy of the selective sphingosine 1-phosphate receptor modulator ozanimod in relapsing multiple sclerosis (RADIANCE): a randomised, placebo-controlled, phase 2 trial

Modulation of sphingosine 1-phosphate (S1P) receptors in a non-selective manner decreases disease activity in patients with multiple sclerosis but has potential safety concerns. We assessed the safety and efficacy of the oral selective S1P receptor modulator ozanimod in patients with relapsing multiple sclerosis. RADIANCE is a combined phase 2/3 trial. Patients with relapsing multiple sclerosis were recruited from 55 academic and private multiple sclerosis clinics in 13 countries across Europe and the USA. Eligible participants were aged 18-55 years, had an Expanded Disability Status Scale (EDSS) score of 0-5·0, and had either one or more relapses in the previous 12 months, or one or more relapses in the past 24 months and one or more gadolinium-enhancing lesions on MRI in the previous 12 months before screening. Participants were assigned by a computer-generated randomisation sequence in a 1:1:1 ratio to ozanimod (0·5 mg or 1 mg) or matching placebo once daily for 24 weeks by an independent, unmasked, statistical team. Trial participants, study site personnel, MRI assessors, steering committee members, and the study statistician were masked to treatment assignment. To attenuate first-dose cardiac effects, ozanimod was up-titrated from 0·25 mg to 0·5 mg or 1 mg over 8 days. The primary endpoint was the cumulative number of total gadolinium-enhancing MRI lesions measured by an independent MRI analysis centre at weeks 12-24 after treatment initiation. Analysis was by intention to treat. Here, we report results from the 24-week phase 2 trial. This trial is registered with ClinicalTrials.gov, number NCT01628393. The 2-year phase 3 trial is ongoing. The first patient was randomised on Oct 18, 2012, and the final visit of the last randomised patient was on May 11, 2014. The intention-to-treat and safety population consisted of 258 participants, 88 were assigned placebo, 87 ozanimod 0·5 mg, and 83 ozanimod 1 mg; 252 (98%) patients completed the assigned treatment. The mean cumulative number of gadolinium-enhancing lesions at weeks 12-24 was 11·1 (SD 29·9) with placebo compared with 1·5 (3·7) with ozanimod 0·5 mg (odds ratio 0·16, 95% CI 0·08-0·30; p<0·0001) and 1·5 (3·4) with ozanimod 1 mg (odds ratio 0·11, 95% CI 0·06-0·21; p<0·0001). Three serious adverse events unrelated to treatment were reported in patients assigned ozanimod 0·5 mg: optic neuritis, somatoform autonomic dysfunction, and cervical squamous metaplasia (HPV-related). No serious infectious or cardiac adverse events were reported, and no cases of macular oedema arose. The most common adverse events in the ozanimod 0·5 mg and 1 mg groups compared with placebo were nasopharyngitis (11 and five vs 12), headache (five and three vs eight), and urinary-tract infections (six and two vs two). The maximum reduction in mean heart rate by Holter monitoring during the first 6 h in ozanimod-treated participants was less than 2 beats per min (bpm) compared with baseline, with no patient having a minimum hourly heart rate less than 45 bpm. Electrocardiograms and 24-h Holter monitoring showed no increased incidence of atrioventricular block or sinus pause with ozanimod. Ozanimod significantly reduced MRI lesion activity in participants with relapsing multiple sclerosis, with a favourable safety profile over a period of 24 weeks. These findings warrant phase 3 trials, which are ongoing. Receptos, Inc.