新型免疫抑制剂2-氨基-2-[2-(4-正辛基苯基)乙基]-1,3-丙二醇盐酸盐(FTY720)的合成

目的 合成新型免疫抑制剂2-氨基- 2-[2-(4-正辛基苯基)乙基]-1,3-丙二醇盐酸盐(FTY720)。方法 以氯化苄和溴代正庚烷为起始原料,经Wurtz偶联、氯乙酰化、缩合、羰基还原、酯基还原、去乙酰化和成盐反应得到目的化合物FTY720,总收率为22.5％。结果与结论 目标化合物的结构经 1H-NMR、13C-NMR和MS确证。中间体的1H-NMR谱和熔点与文献值相符。该合成路线成本低廉,步骤较短,操作简单。     

2-正丁基-5-甲酰基咪唑的合成

目的合成抗高血压药伊普沙坦的关键中间体2-正丁基-5-甲酰基咪唑。方法乙二胺与戊腈在硫粉的作用下环合得到2-正丁基咪唑啉（2），经脱氢、碘化、还原、羟甲基化、氧化、催化氢解得到目标化合物2-正丁基-5-甲酰基咪唑（1）。结果与结论目标化合物的结构经MS、^1H-NMR谱确证，优化后的合成路线，所用试剂易得、操作简单、反应条件温和，总收率为10．8％。     

1-（2m硝基-1-咪唑基）-2-O-四氢吡喃基-3-O-甲苯磺酰基丙二醇的合成

且的合成乏氧显像剂FMISO的前体1-（2-硝基-1-咪唑基）-2-O-四氢吡喃基-3-O-甲苯磺酰基丙二醇（NITTP）。方法以甘油为原料，通过酯化、羟基保护以及亲核取代反应合成目标化合物。结果与结论目标化合物的总收率为29％，结构经^1H-NMR、^13C-NMR、MS确证，本法生产成本低，收率较高。     

2-（E）-亚苄基-5-芳氨基甲基环戊醇类化合物的合成及其抗炎活性研究

目的设计合成2-（E）-亚苄基-5-芳氨基甲基环戊醇类化合物，并对其抗炎活性进行初步的评价。方法以环戊酮为起始原料，通过Stork烯胺反应、Mannich缩合反应、胺交换反应和选择性还原制备目标化合物；以二甲苯致小鼠耳肿胀模型测试目标化合物的抗炎活性。结果共合成16个新化合物，其结构经^1H-NMR和MS谱确证。结论初步药理实验结果显示，4个目标化合物具有较强的抗炎活性。     

3-氟-4-(2-芳基噻唑-4-基)苯基(噁)唑烷酮类化合物的合成及抗菌活性

目的设计并合成3-氟-4-（2-芳基噻唑-4-基）苯基噁唑烷酮类化合物，初步评价其体外抗菌活性。方法 以（R）-环氧氯丙烷和间氟苯基异氰酸酯为起始原料，通过新的合成路线制备了目标化合物；采用微量液体稀释法，测定目标化合物的体外抗菌活性。结果与结论 合成了14个新化合物。其结构经^1H-NMR、MS确认，10个化合物显示出不同程度的抗菌活性，化合物IXc和Xc的活性较好，有进一步研究的价值。     

3-(2-烷氧基-5-甲基苯基)-3-苯基丙胺类化合物的合成

目的 设计合成3－（2－烷氧基－5－甲基苯基）－3－苯基丙胺类化合物，从中筛选有抗尿失禁作用而无口干等不良反应的活性化合物。方法 以肉桂酸和对甲酚为起始原料，经环合、甲基化、酰胺化、还原或再经脱甲基化得目标产物。结果 共合成10个新化合物，经IR，^1H-NMR等确证其结构。     

1，5-二芳基吡唑衍生物的合成及其对环氧合酶2的抑制活性

目的设计合成一系列1，5-二芳基吡唑衍生物，并考察其对环氧合酶2的抑制活性。方法以苯乙酮或对甲基苯乙酮为原料，经过缩合、环合、水解、还原、酯化等多步反应，得到1，5-二芳基．3-取代吡唑衍生物。结果与结论共合成了10个中间体和10个目标化合物，其中，18个化合物（8个中间体和10个目标化合物）是未见文献报道的新化合物，其结构经MS和^1H-NMR确证。所有目标化合物均具有一定的环氧合酶2抑制活性，其中化合物1i的抑制率为31．77％。     

2-乙酰氧基-5-乙酰氨基苯甲酸的合成及镇痛抗炎作用

目的：合成2-乙酰氧基-2-乙酰氨基苯甲酸并进行镇痛、抗炎作用的初步研究。方法：以水杨酸和苯胺为原料，经重氮、偶合、还原及酰化合成目标化合物，化学结构经元素分析、IR和^1H-NMR确定；采用小鼠醋酸扭体、耳部肿胀法研究镇痛和抗炎作用。结果：目标化合物的熔点为170-173℃，中和法测量99.2％，收率42.7％（以水杨酸计）。小鼠灌胃后抑制醋酸所致疼痛的ED50为1.2mmol/kg，抑制二甲苯所致耳廓炎症的ED50 为2.3mmol/kg，LD50为2598mg/kg。结论：2-乙酰氧基-5-乙酰氨基苯甲酸合成简单，收率稳定，具有比阿司匹林和对乙酰氨基酚更强的镇痛和抗炎作用。     

2-甲基-5-(E)-(邻甲氧基苯亚甲基)环戊酮Mannich碱类化合物的合成及其抗炎活性

目的 设计合成2-甲基-5-（E）-（邻甲氧基苯亚甲基）环戊酮曼尼希碱类化合物，并对其抗炎活性进行初步评价。方法 以N-环戊烯基吗啉、邻甲氧基苯甲醛及多种胺类为原料，经多步反应全成目标化合物，并以二甲苯致小鼠耳肿胀模型测试目标化合物的抗炎活性。结果 共合成8个新化合物，经IR，^1H-NMR和MS确证其结构。其中两个化合物的抗炎活性与阿司匹林相当。结论 羧基a位被烷基取代的环戊酮曼尼希碱仍具有较强的抗炎作用。     

(E)-5-(2-溴乙烯基)-1-β-D-阿拉伯呋喃糖尿嘧啶的合成

目的研究（E）-5-（2-溴乙烯基）-1-β-D-阿拉伯呋喃糖尿嘧啶的合成。方法以四乙酰核糖为原料与2，4-二甲硅烷基尿嘧啶偶联得2’，3’，5＇-三-O-乙酰核糖尿嘧啶（2），经醇解、脱水环合、开环得1-（β-D-阿拉伯呋喃糖）尿嘧啶（5），碘化得化合物6，再与丙烯酸甲酯偶联得化合物7，经碱水解得8，最后经Hunsdfiecker反应合成目标化合物1。结果与结论目标化合物1及各产物结构经MS、1H-NMR、元素分析等确证。该工艺路线原料易得，反应无α-异构体副产物生成，无需色谱分离操作，可用于规模制备。     

FTY-720通过TGF-β1/P38 MAPK/NF-κB信号通路对小鼠肺纤维化模型的影响

目的探讨FTY-720对博来霉素诱导的小鼠肺纤维化的影响及其作用机制。方法将40只无特定病原体级健康雄性Balb/c小鼠随机分为对照组、博来霉素组、生理盐水+FTY-720对照组、博来霉素+FTY-720组,每组各5只,在7天、14天处死小鼠,共8组。应用HE染色和Masson染色观察肺组织炎症及肺纤维化情况;通过瑞士-吉姆萨染色计数支气管肺泡灌洗液细胞;BCA法测定BALF蛋白含量;ELISA方法检测BALF上清液中炎症因子IL-1β、TNF-α的表达水平;免疫组织化学法检测肺组织中COL1A1的表达水平;Western blot方法检测TGF-β1、p-P38 MAPK和NF-κB表达水平的变化。结果FTY-720显著降低BLM诱导增加的小鼠肺组织细胞外胶原沉积;减少小鼠肺组织BALF中IL-1β和TNF-α表达水平,和蛋白质含量、细胞数;FTY-720抑制TGF-β1活性与P38 MAPK的磷酸化,进而抑制NF-κB的表达与活化。结论FTY-720通过抑制TGF-β1/P38 MAPK/NF-κB信号通路,进而抑制博来霉素诱导小鼠的肺纤维化。     

Sphingosine and its analog, the immunosuppressant 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol, interact with the CB1 cannabinoid receptor

Sphingosine-1-phosphate (S1P) and cannabinoid receptors are G-protein-coupled receptors that mediate the effects of S1P and endocannabinoids, respectively. Cannabinoid receptors also mediate the effects of Delta9-tetrahydrocannabinol, the primary psychoactive ingredient in marijuana, whereas S1P receptors contribute to the immunosuppressant effects of 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol (FTY720). FTY720 is a sphingosine analog that can prevent renal graft rejections and suppress a variety of autoimmune disorders in animal models and clinical trials. We now report that both FTY720 and sphingosine interact with CB1 but not CB2 cannabinoid receptors. FTY720 and sphingosine inhibited the binding of the CB1-selective antagonist [3H]N-(piperidinyl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide ([3H]SR141716A) and the cannabinoid agonist [3H](-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclohexanol ([3H]CP55,940) in a concentration-dependent manner in both CB1-expressing cell lines and mouse cerebellum. However, these compounds did not significantly alter [3H]CP55,940 binding to CB2 receptors. In G-protein activation assays, FTY720 and sphingosine inhibited the maximal stimulation of guanosine 5'-O-(3-[35S]thio)triphosphate binding by the cannabinoid agonist R-(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-naphthalenyl)methanone mesylate (WIN55,212-2) in a concentration-dependent manner, and this antagonist effect was not mimicked by S1P. FTY720 and sphingosine also inhibited activation of extracellular signal-regulated kinases 1 and 2 and Akt by WIN55,212-2 in intact Chinese hamster ovary (CHO) cells expressing CB1 receptors and attenuated WIN55,212-2-stimulated internalization of a fluorescence-tagged CB1 receptor in CHO cells. Moreover, both FTY720 and sphingosine produced rightward shifts in the concentration-effect curves of cannabinoid agonists for G-protein activation, indicating that they act as competitive CB1 antagonists. These results suggest that the CB1 receptor could be a novel target of FTY720 and that sphingosine could be an endogenous CB11 antagonist.     

Discovery of Clinical Candidate GSK1842799 As a Selective S1P1 Receptor Agonist (Prodrug) for Multiple Sclerosis

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Intercellular Lipid Mediators and GPCR Drug Discovery

G-protein-coupled receptors (GPCR) are the largest superfamily of receptors responsible for signaling between cells and tissues, and because they play important physiological roles in homeostasis, they are major drug targets. New technologies have been developed for the identification of new ligands, new GPCR functions, and for drug discovery purposes. In particular, intercellular lipid mediators, such as, lysophosphatidic acid and sphingosine 1-phosphate have attracted much attention for drug discovery and this has resulted in the development of fingolimod (FTY-720) and AM095. The discovery of new intercellular lipid mediators and their GPCRs are discussed from the perspective of drug development. Lipid GPCRs for lysophospholipids, including lysophosphatidylserine, lysophosphatidylinositol, lysophosphatidylcholine, free fatty acids, fatty acid derivatives, and other lipid mediators are reviewed.     

The utility of pharmacokinetic-pharmacodynamic modeling in the discovery and optimization of selective S1P(1) agonists

Sphingosine-1-phosphate (S1P(1)) receptor agonists such as Fingolimod (FTY-720) are a novel class of immunomodulators that have clinical utility in the treatment of remitting relapsing multiples sclerosis. This class of compound act by inducing peripheral lymphopenia. Using an integrated pharmacokinetic/pharmacodynamic (PK-PD) approach based on an in vivo rat model, novel S1P(1) agonists were identified with a predicted more rapid rate of reversibility of lymphocyte reduction in human compared to Fingolimod. The in vivo potency of 15 compounds based on PK-PD modelling of the rat lymphocyte reduction model was correlated with in vitro measures of potency at the S1P(1) receptor using β arrestin recruitment and G-protein signalling. A structurally novel S1P(1) agonist was identified and predictions of human pharmacokinetics and clinical dose are presented.     

T cell selective apoptosis by a novel immunosuppressant,FTY720, is closely regulated with Bcl-2

1. A novel immunosuppressant FTY720 caused a significant decrease in peripheral T lymphocytes, but not in B lymphocytes upon oral administration. This decrease was mainly a result of FTY720-induced apoptosis. In this study, we confirmed FTY720-induced T cell selective apoptosis using lymphoma cell lines in vitro. 2. Viability loss, DNA fragmentation, Annexin V binding, and caspases activation (caspase-3, -8, and -9) were observed in Jurkat cells (T lymphoma cells), but not significantly in BALL-1 cells (B lymphoma cells). These results indicated that FTY720 selectively induced apoptosis in T cell lymphoma to a greater extent than in B cell lymphoma, a finding that is similar to the result observed when FTY720 was treated with T lymphocytes and B lymphocytes in vitro. 3. FTY720 released cytochrome c from mitochondria in Jurkat intact cells as well as from isolated Jurkat mitochondria directly, but not from mitochondria in BALL-1 cells nor from isolated BALL-1 mitochondria. 4. BALL-1 cells and B cells had more abundant mitochondria-localized anti-apoptotic protein Bcl-2 than did Jurkat cells and T cells. 5. FTY720-induced apoptosis is inhibited by the overexpression of Bcl-2, suggesting that the cellular Bcl-2 level regulates the sensitivity to FTY720.     

Promising new agents in the prevention of transplant rejection

Promising immunosuppressive drugs designed to prevent rejection have been developed recently. Two monoclonal antibodies directed against the interleukin-2 (IL-2) receptor, daclizumab and basiliximab, have been shown to significantly reduce the incidence of acute rejection without increasing adverse events. Sirolimus (rapamycin), an agent that inhibits T- and B-response at a later stage than cyclosporin, has been shown to be synergistic with cyclosporin in experimental and clinical studies. Ongoing clinical trials have reported that in renal transplantation high doses of sirolimus are as effective as cyclosporin. SDZ-RAD, a derivative of sirolimus, is also under investigation. FTY-720 another promising drug, prolonged the survival of allografts and synergised with cyclosporin and sirolimus in experimental models. Gusperimus (deoxyspergualin), which inhibits IL-1 synthesis, was useful in reversing early and late acute rejection in clinical trials. Antisense oligonucleotides which interfere with intercellular adhesion molecules which are important in rejection, gave encouraging results in primate renal allografts. The availability of these new drugs will be able to further abate the risk of rejection in organ transplantation. However, caution is warranted with their use in order to avoid the risks of over immunosuppression. Today excellent results can be obtained with the available drugs. Newer immunosuppressive schedules should be designed, not only to reduce the risk of rejection, but also to obtain a better therapeutic index that allows a further improvement of the graft survival while minimising the comorbidity and drug-related toxicity.     

Effect of a synthetic prostaglandin E2 analogue,RS-86505-007, on plasma lipids and lipoproteins in patients with moderate hypercholesterolaemia: efficacy and tolerance of treatment and response in different apolipoprotein polymorphism groups

A double-blind, placebo-controlled ascending dose trial was carried out to evaluate the hypocholesterolaemic efficacy and tolerance of RS-86505-007, a prostaglandin E₂ analogue, in moderately hypercholesterolaemic patients. Twenty-four patients received an oral dose of RS-86505-007 3 g t.i.d. and a separate group of 26 patients 6 g t.i.d. for 6 weeks. Plasma total and low-density lipoprotein (LDL) cholesterol concentrations decreased after 2 weeks of treatment, and the reductions were dose dependent. After 6 weeks of treatment (6 g t.i.d.), the reductions from baseline in total and LDL cholesterol concentration were 14.6% and 18.5%, respectively. No changes in the plasma concentration of triglycerides or high-density lipoprotein (HDL) cholesterol were observed. RS-86505-007 tended to reduce total and LDL cholesterol concentrations less in patients with the 4 allele of the apolipoprotein E than in those with 3 allele. In contrast, the XbaI or EcoRI polymorphisms of the apolipoprotein B gene seemed to have no effect on the hypocholesterolaemic efficacy of the drug. The drug had no effect on the lipoprotein (a) concentration. Sixty-three per cent of patients receiving 3 g t.i.d. and 81% receiving 6 g t.i.d. had adverse events, two-thirds of which related to the gastrointestinal tract. One patient in the 3-g group and three patients in the 6-g group terminated the study prematurely due to adverse effects. In conclusion, a prostaglandin E₂ analogue, RS-86505-007, has a potent cholesterol-lowering effect, but its grastrointestinal adverse effects are marked. RS-86505-007 could, however, be used as a tool in mechanistic studies of cholesterol metabolism.     

Effect of pretreatment with FTY720 and cyclosporin on ischaemia-reperfusion injury of the liver in rats

The effect of pretreatment with FTY720 (2-amino-2-[2-(4-octylphenyl)ethyl]-1,3-propanediol hydrochloride) or cyclosporin, or both, on neutrophil-mediated injury has been examined by use of a rat model of transient clamping of hepatic flow. Pretreatment with FTY720 alone or with cyclosporin induced a marked reduction of circulatory lymphocytes, whereas the use of these drugs in combination was very effective at suppressing the elevation of the number of peripheral polymorphonuclear neutrophils (PMN) after reperfusion. Pretreatment with cyclosporin, with or without FTY720, significantly reduced hepatic damage, whereas FTY720 alone tended to prolong hepatic damage. Pretreatment of cyclosporin alone, but not in combination with FTY720, significantly reduced the accumulation of PMN and led to lower myeloperoxidase levels in the damaged liver. In conclusion, pretreatment with cyclosporin, with or without FTY720, reduced hepatic damage after warm ischaemia-reperfusion, whereas pretreatment with FTY720 alone tended to prolong this damage.