咖啡因5种主要代谢物的反相高效液相色谱法测定

为保证应用咖啡因代谢探针的正确性,建立了反相高效液相色谱测定尿中咖啡因代谢物的方法。采用日本岛津 Shim Pack CLC-ODS柱(5μm),以甲醇—乙腈—0.05%醋酸(12∶1∶87)为流动相,咖啡因和它的13种代谢物及内标均能良好分离。重点对其中5个主要代谢物AFMU,1U,1X,17U,17X进行了测定。结果表明这5种代谢物回收率均在87%以上,日内和日间误差均小于3%,显示了方法的稳定性。并对120例志愿者尿中5种主要代谢物浓度进行了测定,为进一步评价多种药物代谢酶活性创造条件。     

21-氯代达哪唑及其C_6-脱氢物的合成

达哪唑(danazol)国产品名炔睾醇(Ⅲa),是治疗子宫内膜异位症的药物,并具有多方面的生理活性,既能抑制垂体前叶腺素的合成与释放,也是3β-羟基甾体脱氢酶(3βHSD)的抑制剂,国外曾作为女用口服避孕药或与睾丸素酯合用作男性抗生育剂,近年来还作为女用事后避孕药。为了寻找新的抗生育调节剂以适应计划生育工作的需要,我们对达哪唑进行结构改造研究,合成了它的21-氯化物(Ⅳa)及其C_6-脱氢物(Ⅳb)。其合成路线如下:     

小檗碱对局灶性脑缺血大鼠血小板聚集及血浆TXB2和6-keto-PGF1a水平的影响

以大鼠可逆性大脑中动脉梗塞(MCAO)致局灶性脑缺血为模型,观察小檗碱对大鼠MCAO24h后血小板粘附、聚集、血栓形成及血浆TXB2和PGI₂生成的影响。结果表明,小檗碱20mg·kg^-1·d^-1ipl,3或5d,明显降低MCAo24h后血小板粘附性及ADP、胶原和花生四烯酸诱导的血小板聚集率,抑制血浆TXB₂水平。同剂量ip3或5d,则抑制血栓形成。提示小檗碱可能通过其抗血小板粘附和聚集及影响花生四烯酸代谢而发挥抗脑缺血作用。     

血浆中去甲地西泮及代谢物奥沙西泮的HPLC测定方法和大鼠口服药代动力学

采用高效液相色谱法测定去甲地西泮(去甲安定)及其代谢产物奥沙西泮。以RP-C₁₈为固定相,乙腈—0.01mol·L^-1醋酸钠(pH3.8,33.3:66.6)为流动相,地西泮为内标物,紫外波长240nm处定量测定。去甲地西泮、奥沙西泮和内标物的保留时间分别为2.8min,4.85min和8.5min;绝对回收率分别为74%,86%和86%。奥沙西泮在35.3～2260ng·ml^-1,去甲地西泮在20～2560ng·ml^-1血浆浓度范围内线性关系良好,r=0.9997和r=0.9998。二药的最低检测浓度分别为10ng·ml、和7ng·ml^-1;日内和日间相对标准偏差(RSD)均分别小于6%和10%(n=5)。多种常用药物对样品的色谱峰无干扰。并用此法研究了大鼠单次口服去甲地西泮的药代动力学。     

尿中麻黄碱类药物的气相色谱检测法

麻黄碱及其类似物有兴奋作用,属体育比赛禁用药。这类化合物的化学结构相似,故有相似的鉴别特征。本文报道了以气相色谱对尿中麻黄碱(ephedrine)、甲基麻黄碱(methylephedrine)、乙基麻黄碱(ethylephedrine)、去甲麻黄碱(norephedrine)及去甲伪麻黄碱(cathine)进行检测,最后以气质联用法进行确证。本法操作简便,灵敏度高,结果可靠。     

A novel role for the metabotropic glutamate receptor-7: modulation of faecal water content and colonic electrolyte transport in the mouse

Background and purpose:

Increasing evidence implicates metabotropic glutamate receptor mGlu₇ in the pathophysiology of stress-related disorders such as depression and anxiety. Mood disorders are frequently associated with gastrointestinal (GI) dysfunction; however, the role of mGlu₇ receptors outside the CNS is unknown. This present study investigated the expression and possible functional role of mGlu₇ receptors in the mouse colon.

Experimental approach:

Expression of mGlu₇ receptor mRNA and protein was studied in mouse colon by in situ hybridization and Western blotting. Effects of the selective mGlu₇ receptor agonist AMN082 on defecation and faecal parameters were studied in an isolation-induced stress model. AMN082 effects on ion transport and neuronal intracellular signalling were examined via Ussing chambers and calcium imaging.

Key results:

mGlu₇ receptor mRNA and protein were highly expressed in colon mucosa. Stress-induced faecal output was unaffected by AMN082, although faecal water content was increased. In mucosa/submucosa preparations, 100 nM and 1 µM AMN082 increased bethanechol-induced changes in short-circuit current in the Ussing chamber. This was sensitive to tetrodotoxin. Also, 100 nM AMN082 significantly increased calcium signalling in a subset of submucosal neurons.

Conclusions and implications:

Activating mGlu₇ receptors increased colonic secretory function in vivo and ex vivo. In a group of submucosal neurons, AMN082 strongly induced calcium signalling and the presence of submucosal nerves was required for the AMN082-dependent increase in secretion. These data suggest that targeting mGlu₇ receptors may be useful in the treatment of central components of stress disorders and also stress-associated GI dysfunction such as diarrhoea or constipation.     

圆二色谱在天然产物——二萜双向酯和娃儿藤碱的结构分析中的应用

本文报道了四个二萜双内酯和五个娃儿藤碱的圆二色谱、扇形图和反八区律,其中6和8为新化合物。文中提出的反八区律图首先应用于二萜双内酯的构型测定,我们应用有关的规则,所预测的这些化合物的Cotton效应和实测结果一致,从而指定了它们的绝对构型。     

A novel, orally active LPA1 receptor antagonist inhibits lung fibrosis in the mouse bleomycin model

Background and purpose:

The aim of this study was to assess the potential of an antagonist selective for the lysophosphatidic acid receptor, LPA₁, in treating lung fibrosis We evaluated the in vitro and in vivo pharmacological properties of the high affinity, selective, oral LPA₁-antagonist (4′-{4-[(R)-1-(2-chloro-phenyl)-ethoxycarbonylamino]-3-methyl-isoxazol-5-yl}-biphenyl-4-yl)-acetic acid (AM966).

Experimental approach:

The potency and selectivity of AM966 for LPA₁ receptors was determined in vitro by calcium flux and cell chemotaxis assays using recombinant and native cell cultures. The in vivo efficacy of AM966 to reduce tissue injury, vascular leakage, inflammation and fibrosis was assessed at several time points in the mouse bleomycin model.

Key results:

AM966 was a potent antagonist of LPA₁ receptors, with selectivity for this receptor over the other LPA receptors. In vitro, AM966 inhibited LPA-stimulated intracellular calcium release (IC₅₀ = 17 nM) from Chinese hamster ovary cells stably expressing human LPA₁ receptors and inhibited LPA-induced chemotaxis (IC₅₀ = 181 nM) of human IMR-90 lung fibroblasts expressing LPA₁ receptors. AM966 demonstrated a good pharmacokinetic profile following oral dosing in mice. In the mouse, AM966 reduced lung injury, vascular leakage, inflammation and fibrosis at multiple time points following intratracheal bleomycin instillation. AM966 also decreased lactate dehydrogenase activity and tissue inhibitor of metalloproteinase-1, transforming growth factor β1, hyaluronan and matrix metalloproteinase-7, in bronchoalveolar lavage fluid.

Conclusions and implications:

These findings demonstrate that AM966 is a potent, selective, orally bioavailable LPA₁ receptor antagonist that may be beneficial in treating lung injury and fibrosis, as well as other diseases that are characterized by pathological inflammation, oedema and fibrosis.     

丁基苯酞对局部脑缺血大鼠神经元迟发性损伤及细胞内钙的影响

观察了丁基苯酞(NBP)对局部脑缺血迟发性脑梗塞和神经功能缺损的影响。结果表明,大鼠大脑中动脉阻断(MCAO)后2h分别poNBP80,160和240mg·kg^-1,均能明显降低脑梗塞面积,抑制率分别为49.0%,69.5%及85.1%,并明显改善神经功能缺失。在MCAO前1h予防给NBP(160mg·kg^-1po),也可明显缩小脑梗塞面积和改善神经功能缺失;在MCAO前连续口服7d,NBP剂量为80mg·kg^-1·d^-1,对上述指标均有改善作用。NBP对高K⁺引起大鼠脑突触体内Ca²⁺含量升高无影响。以上结果提示NBP对迟发性神经元损伤有保护作用。