尼美舒利胶囊人体相对生物利用度研究

采用双交叉试验设计,用ＨＰＬＣ－ＵＶ法测定不同时相的血药浓度,进行药代动力学分析,结果表明：１０名健康志愿者分别一次口服受试制剂及参比片剂 ２００ ｍｇ后所得的药时曲线均符合一室开放模型。尼美舒利胶囊与片剂的主要药动学参数分别为：Ｔ１／２Ｋ（ｈ）,４．２５±０．５８和４ ６７±０．４５;Ｃｍａｘ（ｍｇ．Ｌ－１）,９２８±２２０和８．９２±２．４８;Ｔｍａｘ（ｈ）,２．８８±０．５１和２．７９±０．５５;ＭＲＴ（ｈ）,９．３０±０．９２和９５０±０６０;ＡＵＣ０－２４（ｍｇ·ｈ·Ｌ－１）,８７．０２±２６．８８和８６．９８±２５．３３;这些参数经方差分析法证明无显著性差异（Ｐ＜０．０５）。尼美舒利胶囊对片剂的相对生物利用度（Ｆ％）为：１００·５４±１１．０１。两者的ＡＵＣ、Ｃｍａｘ、Ｔｍａｘ 及ｌｎＡＵＣ、ｌｎＣｍａｘ、ｌｎＴｍａｘ经交叉试验下的方差分析法证明无显著性差异,经双单侧检验证明尼美舒利胶囊与片剂具有生物等效性。     

非甾体抗炎药尼美舒利的合成及结构鉴定

报告了非甾体抗炎药尼美舒利的合成工艺及结构鉴定。工艺过程包括威廉生反应、还原反应、磺酰化反应和硝化反应,总收率５５％,产品的含量≥９９．０％,通过元素分析、紫外吸收光谱、红外吸收光谱、~１Ｈ－核磁共振谱、~（１３）Ｃ－核磁共振谱和质谱确证了结构。     

Influence of piperine on nimesulide induced antinociception

Piperine (1-peperoyl piperidine), an alkaloid extracted from Piper nigrum Linn is an inhibitor of hepatic and other enzymes involved in the biotransformation of drugs. In the present study piperine showed a dose dependent synergistic effect on nimesulide induced antinociception, in the acetic acid induced writhing test in mice. Piperine at a dose of 10 mg/kg significantly (p < 0.001) increased the analgesic activity of nimesulide administered at a submaximal dose of 6.5 mg/kg. In the formalin test, nimesulide alone (10 mg/kg, oral) did not modify phase I or nociceptor mediated pain while a combination of nimesulide (10 mg/kg) with piperine (10 mg/kg) significantly decreased it. In phase II or inflammatory pain, duration of formalin induced behaviour was 80 ± 7 s, 61 ± 7.3 s and 5.33 ± 3.3 s in control, nimesulide treated and piperine plus nimesulide treated groups respectively, indicating a synergistic activity of piperine with nimesulide. The antinociceptive effect correlated well with increased plasma concentration of nimesulide. The plasma concentration after oral administration of nimesulide (10 mg/kg) alone was 8.03 ± 0.99 ug/mL. However, when it was administered with piperine (10 mg/kg), the plasma concentration of nimesulide increased to 11.9 ± 0.23 ug/mL. This indicates that piperine inhibits the biotransformation and metabolism of nimesulide leading to significantly (p < 0.05) higher levels of drug in the systemic circulation. The findings of the present study suggest that piperine could be used as a biological enhancer when coadministered with nimesulide. © 1998 John Wiley & Sons, Ltd.     

尼美舒利对乳腺癌细胞放射增敏作用的研究

 目的 观察尼美舒利（nimesulide）对人乳腺癌细胞MCF-7辐射敏感性的影响,并探讨其可能机制。方法 实验分为对照组、加药组、单纯照射组(radiation treatment,RT)及加药照射组,用噻唑蓝（MTT）法检测尼美舒利对MCF-7细胞生长抑制率的影响,并选择用药后72 h测得的IC₂₀-IC₃₀（抑制浓度,inhibiting concentration,IC）对应的药物浓度作为辐射增敏工作浓度;克隆形成实验检测尼美舒利对MCF-7的放射增敏作用;通过流式细胞仪检测细胞周期分布的变化。Western印迹法检测与凋亡相关蛋白Bcl-2和Bax的表达水平;并用单细胞凝胶电泳技术检测DNA的损伤及修复。结果 尼美舒利对MCF-7细胞的生长抑制作用呈时间依赖性和剂量依赖性,但对周期分布无明显变化。尼美舒利加药照射组与单纯照射组比较,克隆存活曲线的肩区变窄,bcl-2表达下调,但对Bax表达没有明显影响。尼美舒利本身不明显增加MCF-7的DNA损伤,但可明显延缓放射引起的SSB的修复。结论 提示尼美舒利对人乳腺癌MCF-7细胞株具有辐射增敏作用,其放射增敏机制可能通过抑制DNA损伤修复并下调bcl-2来得以实现。     

Evaluation of guggulipid and nimesulide on production of inflammatory mediators and GFAP expression in LPS stimulated rat astrocytoma,cell line (C6)

Aim of the study

The present study was designed to investigate effect of guggulipid, a drug developed by CDRI and nimesulide on LPS stimulated neuroinflammatory changes in rat astrocytoma cell line (C6).

Materials and methods

Rat astrocytoma cells (C6) were stimulated with LPS (10 μg/ml) alone and in combinations with different concentrations of guggulipid or nimesulide for 24 h of incubation. Nitrite release in culture supernatant, ROS in cells, expressions of COX-2, GFAP and TNF-α in cell lysate were estimated.

Results

LPS (10 μg/ml) stimulated C6 cells to release nitrite, ROS generation, up regulated COX-2 and GFAP expressions at protein level and TNF-α at mRNA level. Both guggulipid and nimesulide significantly attenuated nitrite release, ROS generation and also down regulated expressions of COX-2, GFAP and TNF-α. Guggulipid and nimesulide per se did not have any significant effect on C6 cells.

Conclusion

Results demonstrate the anti-inflammatory effect of guggulipid comparable to nimesulide which suggest potential use of guggulipid in neuroinflammation associated conditions in CNS disorders.     

尼米舒利对人肺腺癌A549细胞株COX-2表达及细胞增殖和凋亡的影响

目的:探讨尼米舒利(NIM)对人肺腺癌A549细胞株环氧化酶-2(COX-2)表达和细胞增殖及凋亡的影响。方法:RT-PCR法、Western blot法及流式细胞术分别检测对照组和NIM干预组(NIM 25 μmol/L、50 μmol/L、100μmol/L、200 μmol/L干预48 h)A549细胞COX-2 mRNA表达、蛋白表达、细胞周期及凋亡率的变化;MTT法检测各组干预24 h、48 h、72 h后A549细胞增殖的抑制率。结果:NIM呈浓度依赖性抑制A549细胞COX-2 mRNA及蛋白表达(r分别为-0.934、-0.923,P均<0.01);NIM可引起细胞周期变化,随着干预浓度的增大,G0/G1期细胞增多,S期细胞减少,G2/M期细胞变化不明显;NIM可促进A549细胞凋亡,其凋亡率呈浓度依赖性(r=0.994,P<0.01);NIM对A549细胞的增殖有抑制作用,随着干预浓度的增大和时间延长,抑制率增加。结论:NIM对A549细胞COX-2表达的抑制可能是细胞增殖受抑、凋亡增加的重要原因。     

HPLC法测定尼美舒利胶囊有关物质

目的HPLC法测定尼美舒利胶囊中的有关物质。方法D iamonsil-C18柱,甲醇-0.1%(ρ)磷酸氢二钠溶液(体积比65∶35)为流动相,流速为1.0 mL/m in,检测波长254 nm,柱温为室温。结果该法能有效地分离胶囊中的尼美舒利及有关物质,各杂质峰面积的和不大于1%对照溶液的尼美舒利主峰面积。结论该方法可测定尼美舒利胶囊的有关物质,方法准确、可靠。     

Nimesulide interaction with membrane model systems: Are membrane physical effects involved in nimesulide mitochondrial toxicity?

Nimesulide (NIM), a widely used nonsteroidal anti-inflammatory drug (NSAID), is known to interfere with mitochondrial physiology and to cause idiosyncratic hepatotoxicity. In this study, we characterized the effects of NIM on the physical properties of membrane models containing the main phospholipid classes of the inner mitochondrial membrane: phosphatidylcholine (PC), phosphatidylethanolamine (PE) and cardiolipin (CL). NIM binding/incorporation was observed with the mitochondrial membrane mimicking model composed of dioleoyl PC (DOPC), dioleoyl PE (DOPE) and tetraoleoyl CL (TOCL) at a 1:1:1 M ratio, as well as an increase of membrane permeability, monitored by calcein release, and an increase of lipid disorder, evaluated by fluorescence anisotropy of DPH-PA. Consistently, DSC thermograms of dipalmitoyl PC (DPPC) and a mixture of dipalmitoyl PE (DPPE) and TOCL (7:3 M ratio) showed a NIM-induced decrease of the cooperativity of the phase transition and a shift of the DPPC endotherm to lower temperatures. On the other hand, ³¹P NMR studies with the ternary lipid model indicated a stabilizing effect of NIM on the lipid bilayer structure. Quenching of the fluorescent probes DPH and DPH-PA revealed a peripheral insertion of NIM in the hydrophobic portion of the bilayer. Our data indicate that NIM may influence mitochondria physiological processes by interfering with membrane structure and dynamics. The relevance of these findings will be discussed in terms of the reported NIM effects on mitochondria transmembrane potential, protonophoresis, and induction of the permeability transition pore.     

尼美舒利颗粒溶出度测定方法研究

目的研究尼美舒利颗粒溶出度。方法以磷酸盐缓冲液(pH=8.8)为溶出介质,采用篮法进行溶出度测定,转速:100 r.min-1,用紫外分光光度法在393 nm波长处测定。结果绘制的溶出曲线表明20 min内尼美舒利颗粒可溶出80%以上,确定的溶出时间30 min,限度为80%。结论测定6批样品的溶出量均符合规定。     

尼美舒利致儿童溶血性贫血

1例21个月男性患儿因上呼吸道感染给予尼美舒利颗粒50 mg分2次服用（间隔15 h）。第1次服药后3 h,患儿出现抽搐、面色苍白、小便变黄。第2次服药后15 min再次出现抽搐,面色苍白进行性加重,小便呈浓茶色。入院后实验室检查：C反应蛋白79.0 mg/L;白细胞计数35.6×109/L,中性粒细胞计数27.1×109/L,血红蛋白33 g/L,血小板计数471×109/L;凝血酶时间17.2 s,凝血酶原时间20.4 s,国际标准化比值1.63,活化部分凝血活酶时间27.5 s,纤维蛋白原2.95 g/L。予心电监护、吸氧,静脉滴注地塞米松、甲泼尼龙及人免疫球蛋白,静滴碳酸氢钠,输注洗涤红细胞。患儿病情加重,出现昏迷、四肢抽动,血压131/103 mm Hg（1 mm Hg=0.133 kPa）,心率153次/min。予血浆置换、抗感染、免疫抑制、水化及营养支持治疗,患儿病情好转。入院第8天复查血常规：白细胞计数7.0×109/L,中性粒细胞计数5.3×109/L,血红蛋白117 g/L,血小板311×109/L,网织红细胞0.02。