Kaplan J M, Badger A M, Ruggieri E V, Olivera D L, Newman-Tarr T, Bugelski P J
Department of Toxicology, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406-0939.
J Clin Lab Immunol. 1991 Dec;36(4):49-58.
SK&F 105685 (N,N-Dimethyl-8,8-dipropyl-2-azaspiro[4.5]decane-2-propanamine+ ++ dihydrochloride) is a novel azaspirane with beneficial activity in animal models of autoimmune diseases such as adjuvant-induced arthritis and experimental autoimmune encephalomyelitis in the Lewis rat and lupus-like disease in the MRL mouse. The effect of SK&F 105685 on the proliferation of rat lymphoid cells was examined in vitro. The compound inhibited the proliferative response of spleen, thymus and lymph node cells to the mitogen concanavalin A (Con A) in a dose-dependent manner but had little or no effect on the mitogenic response of peripheral blood lymphocytes. Although less potent than cyclosporin A, SK&F 105685 was able to inhibit the proliferation of spleen cells stimulated with PMA and ionomycin or the mitogens phytohemagglutinin (PHA), Con A and pokeweed mitogen (PWM). Relatively early event(s) in cell proliferation were affected by SK&F 105685 since delaying addition of the drug by 24 to 48 hours after Con A stimulation of rat spleen cells resulted in reduced levels of suppression. The mode of action of SK&F 105685 appeared to differ from that of cyclosporin A or rapamycin. Unlike cyclosporin A, SK&F 105685 did not affect IL-2 production by Con A-stimulated spleen cells or the IL-2-producing Jurkat cell line, but, like rapamycin, the compound significantly reduced the IL-2-induced proliferation of rat ConA blasts. These results suggest that inhibition of lymphocyte proliferation by SK&F 105685 may require the activity of an intermediate effector cell(s) present in susceptible populations such as cells from the spleen, thymus, lymph nodes and Con A blast preparations but absent or present in low numbers in resistant populations such as peripheral blood cells. Indomethacin and NG-monomethyl-L-arginine (NGMMA), a competitive inhibitor of nitric oxide synthase, were both unable to relieve SK&F 105685-induced suppression of splenic Con A responses thereby ruling out a role for the production of prostaglandins or nitric oxide by macrophages as an intermediate in drug-mediated suppression. In summary, SK&F 105685 was unable to inhibit lymphoproliferative responses by a mechanism distinct from that of cyclosporin A or rapamycin and which appears to involve regulation of cellular interactions rather than a direct effect on responding lymphocytes.
SK&F 105685（N，N - 二甲基 - 8，8 - 二丙基 - 2 - 氮杂螺[4.5]癸烷 - 2 - 丙胺二盐酸盐）是一种新型氮杂螺环化合物，在自身免疫性疾病动物模型中具有有益活性，如佐剂诱导的关节炎、Lewis大鼠实验性自身免疫性脑脊髓炎以及MRL小鼠的狼疮样疾病。在体外研究了SK&F 105685对大鼠淋巴细胞增殖的影响。该化合物以剂量依赖性方式抑制脾细胞、胸腺细胞和淋巴结细胞对丝裂原刀豆球蛋白A（Con A）的增殖反应，但对外周血淋巴细胞的促有丝分裂反应几乎没有影响。虽然效力不如环孢素A，但SK&F 105685能够抑制用佛波酯（PMA）和离子霉素刺激的脾细胞或丝裂原植物血凝素（PHA）、Con A和商陆有丝分裂原（PWM）的增殖。细胞增殖过程中相对早期的事件受到SK&F 105685的影响，因为在Con A刺激大鼠脾细胞后将药物添加延迟24至48小时会导致抑制水平降低。SK&F 105685的作用方式似乎与环孢素A或雷帕霉素不同。与环孢素A不同，SK&F 105685不影响Con A刺激的脾细胞或产生IL - 2的Jurkat细胞系产生IL - 2，但与雷帕霉素一样，该化合物显著降低IL - 2诱导的大鼠ConA母细胞的增殖。这些结果表明，SK&F 105685对淋巴细胞增殖的抑制可能需要易感群体中存在的中间效应细胞的活性，如来自脾、胸腺、淋巴结和Con A母细胞制剂的细胞，但在抗性群体如外周血细胞中不存在或数量很少。吲哚美辛和一氧化氮合酶的竞争性抑制剂NG - 单甲基 - L - 精氨酸（NGMMA）均无法缓解SK&F 105685诱导的脾Con A反应抑制，从而排除巨噬细胞产生前列腺素或一氧化氮作为药物介导抑制的中间环节的作用。总之，SK&F 105685无法通过与环孢素A或雷帕霉素不同的机制抑制淋巴细胞增殖反应，其机制似乎涉及细胞间相互作用的调节，而不是对反应性淋巴细胞的直接作用。
