Department of Chemical Physiology, Scripps Research Institute, La Jolla, CA 92037, USA.
Mol Pharmacol. 2012 Feb;81(2):166-74. doi: 10.1124/mol.111.076109. Epub 2011 Oct 26.
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, S1P(1) 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 MOG(35-55)- 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 S1P(1)-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 S1P(1) mice. S1P(1) 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 S1P(1) 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. S1P(1)-selective agonists that induce reversible lymphopenia while persisting in the CNS may be effective MS treatments.
多发性硬化症（MS）疗法可调节中枢神经系统（CNS）中的 T 细胞自身免疫，但可能会加剧潜伏感染。芬戈莫德是一种非选择性的鞘氨醇-1-磷酸（S1P）受体激动剂，可引起持续的淋巴细胞减少并在中枢神经系统中积累，是一种治疗多发性硬化症的新方法。我们假设，持续的淋巴细胞减少不是必需的，并且选择性、穿透中枢神经系统、外周短作用、S1P（1）激动剂在 MS 小鼠模型中也将显示出完全的疗效。我们使用每天 10mg/kg 的 2-（4-（5-（3,4-二乙氧基苯基）-1,2,4-恶二唑-3-基）-2,3-二氢-1H-茚-1-基氨基）乙醇（CYM-5442）在髓鞘少突胶质细胞糖蛋白 MOG（35-55）诱导的实验性自身免疫性脑脊髓炎（EAE）出现临床症状时开始治疗，评估 CYM-5442、载体或芬戈莫德治疗 12 天的临床评分、CNS 细胞浸润、脱髓鞘和神经胶质增生，在实验结束时测定 CNS 和血浆中的 CYM-5442 水平，并在最后一次注射后 3 和 24 小时测量血液淋巴细胞减少情况，在方案结束时测定血浆细胞因子水平。在活性 EAE 期间和用 CYM-5442 治疗时，使用流式细胞术和 Western 印迹法通过使用天然定位增强型绿色荧光蛋白标记的 S1P（1）小鼠，定量测量神经元和星形胶质细胞上 S1P（1）-增强型绿色荧光蛋白表达的变化。单独的 S1P（1）激动剂和芬戈莫德（整个剂量间隔内最大程度的淋巴细胞减少）一样降低了病理特征，尽管在每个剂量间隔内完全逆转了淋巴细胞减少。CYM-5442 水平在 CNS 中持续存在，但不在血浆中。与载体相比，CYM-5442 治疗可抑制 EAE 中神经元和星形胶质细胞上的 S1P（1）表达，并且药物治疗的小鼠中关键细胞因子（如白细胞介素 17A）的水平也显著降低。诱导可逆性淋巴细胞减少而在中枢神经系统中持续存在的 S1P（1）选择性激动剂可能是有效的 MS 治疗方法。
