液质联用法测定吴茱萸碱、吴茱萸次碱、去氢吴茱萸碱在大鼠脑脊液和脑组织中的分布

吴茱萸碱、吴茱萸次碱、去氢吴茱萸碱是吴茱萸中主要的生物碱类成分.为研究吴茱萸主要生物碱成分在大鼠脑脊液中的代谢和在脑组织中的分布,吴茱萸碱、吴茱萸次碱、去氢吴茱萸碱以1∶1∶1的质量比混合,以15 mg/kg的剂量灌胃给予大鼠.本文建立了同时测定大鼠脑脊液和脑组织中三个生物碱成分的液质联用方法,测定结果显示,三个生物碱...     

左金丸配伍意义的药物代谢动力学分析

目的研究左金丸在SD大鼠体内的药代动力学,为临床应用提供参考。方法将左金丸及黄连、吴茱萸制成水煎液,对SD大鼠进行灌胃,HPLC法测定并计算小檗碱、吴茱萸次碱的药代动力学参数。结果左金丸组的小檗碱的t1/2延长,tmax缩短,Cmax增大,AUC（0-∞）增加,代谢较缓慢;吴茱萸次碱tmax缩短,Cmax增大,AUC（0-∞）增加。结论左金丸中黄连∶吴茱萸为6∶1时,能很好地发挥作用。     

吴茱萸碱和吴茱萸次碱电喷雾-离子阱质谱裂解途径研究

目的:对吴茱萸碱和吴茱萸次碱的电喷雾质谱裂解机制进行研究。方法:在电喷雾离子源,正、负离子模式下,分析吴茱萸碱和吴茱萸次碱的一级和多级质谱图。结果:解析了吴茱萸碱和吴茱萸次碱多级质谱碎片,在正负离子模式下,主要发生Rretro-Diels-Alder(RDA)裂解。结论:吴茱萸碱和吴茱萸次碱的ESI-MS质谱裂解途径可为吲哚类生物碱的成分研究提供数据参考。     

五味子配伍对吴茱萸主成分在大鼠肠道吸收中的影响研究

目的：研究五味子配伍对吴茱萸提取物中吴茱萸碱和吴茱萸次碱两种有效成分肠吸收动力学的影响及可能机制,从肠吸收角度揭示方剂配伍的科学本质.方法：采用大鼠单向灌流模型,考察维拉帕米和五味子提取物对吴茱萸碱和吴茱萸次碱肠吸收的影响,利用HPLC分别测定吴茱萸碱和吴茱萸次碱的量,计算两种生物碱的肠吸收速率常数（Ka）和表观渗透系数（Papp）.结果：五味子和吴茱萸配比为1∶1时,吴茱萸碱和吴茱萸次碱的Ka和Papp显著减少（P＜0.05）;五味子吴茱萸2∶1时,吴茱萸碱和吴茱萸次碱的Ka和Papp要比配比1∶1时显著减少（P＜0.05）.加入P-gp抑制药维拉帕米（VP）与正常组相比,吴茱萸碱和吴茱萸次碱的Ka和Papp未见显著差异.结论：五味子配伍可抑制吴茱萸中吴茱萸碱和吴茱萸次碱的肠吸收.     

左金丸配伍意义的药代动力学研究

目的研究黄连、吴茱萸单用或以其6∶1伍用(左金丸,消化系统中药)在SD大鼠的药代动力学。方法黄连、吴茱萸及左金丸制成水煎液,灌胃SD大鼠,用液质联用法测定小檗碱、吴茱萸次碱的血药浓度,并计算其药代动力学参数。结果给予左金丸后,小檗碱相对生物利用度增加,t1/2延长,tmax缩短但呈缓慢代谢状态。吴茱萸次碱的tmax提前,Cmax提高,相对生物利用度增加。结论临床应用左金丸30mL.kg-1,间隔为每日1次较为合理。     

吴茱萸中4种单体成分致肾细胞毒性的初步研究

目的探讨吴茱萸中成分吴茱萸碱、吴茱萸次碱、吴茱萸内酯和辛弗林在体外对人胚肾细胞(HEK-293)的影响。方法采用MTT法检测吴茱萸碱、吴茱萸次碱、吴茱萸内酯和辛弗林对肾细胞活力的影响;给药后检测肾细胞培养上清液中的功能性指标乳酸脱氢酶(LDH)含量;采用倒置相差显微镜对给药后的细胞形态进行观察。结果 MTT法显示,8.3～33.2 g.mL-1的吴茱萸碱、5～40 g.mL-1的吴茱萸次碱和50～200 g.mL-1的吴茱萸内酯对HEK-293细胞活力有明显的抑制作用(P<0.01 or 0.05),辛弗林对肾细胞活力无明显影响。4.15～33.2 g.mL-1的吴茱萸碱、5～20 g.mL-1的吴茱萸次碱和50～200 g.mL-1的吴茱萸内酯能显著升高肾细胞上清液中的LDH(P<0.01),给予辛弗林后LDH无变化。给予4.15～33.2 g.mL-1的吴茱萸碱和100～200 g.mL-1的吴茱萸内酯后,肾细胞均不同程度的皱缩、减少、甚至死亡,吴茱萸次碱和辛弗林对肾细胞形态无明显影响。结论吴茱萸碱、吴茱萸次碱和吴茱萸内酯对肾细胞均有毒性作用,而辛弗林对肾细胞无毒性作用。     

34种吴茱萸中吴茱萸碱和吴茱萸次碱的含量测定

建立了吴茱萸中吴茱萸碱和吴茱萸次碱的反相高效液相色谱法 ,并用该法测定了 34种不同产地及不同炮制方法吴茱萸中吴茱萸碱和吴茱萸次碱的含量 .采用峰面积内标法定量 .吴茱萸碱和吴茱萸次碱在 0 5～ 2 0g/mL范围内均呈线性 ,相关系数分别为 0 9999、0 9998,平均回收率分别为 96 6 %、96 0 %     

复方参萸胶囊质量标准研究

目的建立复方参萸胶囊的质量标准。方法对制剂中的吴茱萸、丹参采用薄层色谱法进行定性鉴别;采用反相高效液相色谱法对吴茱萸碱和吴茱萸次碱进行含量测定。结果薄层色谱斑点清晰、分离效果好、专属性强。吴茱萸碱在0.010 84～0.216 8μg,吴茱萸次碱在0.009 68～0.193 6μg范围内线性关系良好,平均回收率:吴茱萸碱为100.24%,RSD为0.35%,吴茱萸次碱为99.68%,RSD为0.42%。结论该方法准确,简便,稳定性和重复性好,可用于该制剂的质量控制。     

提取物纯度对吴茱萸碱和吴茱萸次碱溶解度与表观分配系数的影响

分别采用平衡法和摇瓶法测定高、中、低纯度(71.0%,49.2%和25.4)吴茱萸提取物中吴茱萸碱和吴茱萸次碱的溶解度及表观分配系数,采用HPLC法测定两者的浓度.高中低纯度吴茱提取物中吴茱萸碱和吴茱萸次碱均在PEG-400中溶解度最高,分别为8～14mg/ml和13～19mg/ml:异丙醇中的溶解度最低(均小于7mg/ml).吴茱萸碱和吴茱萸次碱在介质中的溶解度依次为PEG-400＞Tween-80＞RH-40＞MCT>无水乙醉=1,2-丙二醇=1,3-丁二醇=异丙醇.两者的表观分配系数的对数值(1gp)为0.6～1.2.吴茱萸提取物纯度越高,吴茱萸碱和吴茱萸次碱的溶解度越低:纯度与表观分配系数无相关性.pH对溶解度和表观分配系数均无明显影响.     

不同生产工艺的戊己丸中吴茱萸碱和吴茱萸次碱含质量比较

目的:建立戊己丸中吴茱萸碱和吴茱萸次碱含质量的高效液相(HPLC)测定方法,并对超微戊己丸和常规戊己丸中吴茱萸碱和吴茱萸次碱的含质量进行比较。方法:采用色谱柱HypersilC18柱(4.6mm×250mm,5μm),流动相:乙腈水(48∶52),流速:1mL/min,检测波长:225nm。结果:吴茱萸碱和吴茱萸次碱的理论塔板数均大于3000,吴茱萸碱回归方程:A吴茱萸碱=107.73ρ-14.331,r=0.9999,线性范围1.02～20.4μg/mL;吴茱萸次碱回归方程:A吴茱萸次碱=15.596ρ-45.09,r=0.9998,线性范围4.4～176μg/mL。超微戊己丸中吴茱萸碱平均回收率为:99.61%,RSD为0.50%,质量分数为(0.096±0.0148)%;吴茱萸次碱平均回收率为94.85%,RSD为1.00%,质量分数为(0.343±0.0467)%。常规戊己丸中吴茱萸碱平均回收率为98.56%,RSD为1.70%,质量分数为(0.81±0.003)%;吴茱萸次碱平均回收率为99.55%,RSD为0.50%,质量分数为(0.161±0.0075)%。结论:超微戊己丸中吴茱萸碱和吴茱萸次碱的质量分数高于常规戊己丸。     

Induction of cytochrome P450s by rutaecarpine and metabolism of rutaecarpine by cytochrome P450s

Rutaecarpine is an alkaloid originally isolated from the unripe fruit of Evodia rutaecarpa. Recently, rutaecarpine has been characterized to have an anti-inflammatory activity through cyclooxygenase-2 inhibition. In the present studies, the effects of rutaecarpine on liver cytochrome P450 s (P450s) and P450 s involved in the metabolism of rutaecarpine were studied in vivo and in vitro, respectively, because the data are crucial in the early development of rutaecarpine as a new drug candidate. Oral administration to male ICR mice of rutaecarpine for 3 consecutive days induced liver P450 1A-, 2B- and 2E1-selective monooxygenase activities. The induction of P450 1A and 2B by rutaecarpine was confirmed by Western immunoblotting. When rutaecarpine was incubated with rat liver microsomes in the presence of an NADPH-generating system, five metabolites were detected by UV and mass spectral analyses. The 3-methylcholanthrene- and phenobarbital-induced microsomes greatly increased the formation of metabolites. Our present results suggest that rutaecarpine might induce P450 1A and 2B in mice, and that P450 1A and 2B might predominantly metabolize rutaecarpine in rat liver microsomes.     

高效液相色谱法测定大鼠血浆中吴茱萸次碱浓度

建立大鼠血浆中吴茱萸次碱的反相高效液相色谱法。以乙腈 水 (5 8∶4 2 )为流动相 ,地西泮为内标 ,在波长 3 4 5nm处检测。吴茱萸次碱浓度在 0 0 2～ 1 6μg/mL呈线性 ,相关系数为 0 999,高中低 3种浓度平均萃取回收率均大于 80 0 %     

Oxidative metabolism of the alkaloid rutaecarpine by human cytochrome P450.

Rutaecarpine is the main active alkaloid of the herbal medicine, Evodia rutaecarpa. To identify the major human cytochrome P450 (P450) participating in rutaecarpine oxidative metabolism, human liver microsomes and bacteria-expressed recombinant human P450 were studied. In liver microsomes, rutaecarpine was oxidized to 10-, 11-, 12-, and 3-hydroxyrutaecarpine. Microsomal 10- and 3-hydroxylation activities were strongly inhibited by ketoconazole. The 11- and 12-hydroxylation activities were inhibited by alpha-naphthoflavone, quinidine, and ketoconazole. These results indicated that multiple hepatic P450s including CYP1A2, CYP2D6, and CYP3A4 participate in rutaecarpine hydroxylations. Among recombinant P450s, CYP1A1 had the highest rutaecarpine hydroxylation activity. Decreased metabolite formation at high substrate concentration indicated that there was substrate inhibition of CYP1A1- and CYP1A2-catalyzed hydroxylations. CYP1A1-catalyzed rutaecarpine hydroxylations had V(max) values of 1,388 to approximately 1,893 pmol/min/nmol P450, K(m) values of 4.1 to approximately 9.5 microM, and K(i) values of 45 to approximately 103 microM. These results indicated that more than one molecule of rutaecarpine is accessible to the CYP1A active site. The major metabolite 10-hydroxyrutaecarpine decreased CYP1A1, CYP1A2, and CYP1B1 activities with respective IC(50) values of 2.56 +/- 0.04, 2.57 +/- 0.11, and 0.09 +/- 0.01 microM, suggesting that product inhibition might occur during rutaecarpine hydroxylation. The metabolite profile and kinetic properties of rutaecarpine hydroxylation by human P450s provide important information relevant to the clinical application of rutaecarpine and E. rutaecarpa.     

Solid dispersion of rutaecarpine improved its antihypertensive effect in spontaneously hypertensive rats

It was reported previously that rutaecarpine produced a hypotensive effect in phenol‐induced and 2‐kidney, 1‐clip hypertensive rats. However, the same dose of crude rutaecarpine did not produce significant hypotensive effects when applied to spontaneously hypertensive rats (SHR). In the present study, a different dose of rutaecarpine solid dispersion was administered intragastrically to SHR. The systolic blood pressure was monitored by the tail‐cuff method with an electro‐sphygmomanometer. The plasma concentration of rutaecarpine, calcitonin gene‐related peptide (CGRP) and the mRNA levels of CGRP in dorsal root ganglion were determined. The results showed that administration of the solid dispersion significantly increased the blood concentration of rutaecarpine, accompanied by significant hypotensive effects in SHR in a dose‐dependent manner. The levels of plasma CGRP were also elevated significantly, concomitantly with the increased mRNA levels in the dorsal root ganglion in a dose‐dependent manner. It was concluded that a change of the dosage from the crude drug to solid dispersion could improve significantly the efficiency of rutaecarpine absorption and increase its plasma concentration. The anti‐hypertensive effect exerted by rutaecarpine solid dispersion in SHR is mediated by CGRP. Copyright © 2008 John Wiley & Sons, Ltd.     

Rutaecarpine衍生物的合成及体外抗血小板聚集活性

目的合成新的rutaecarpine衍生物并对其抗血小板聚集活性进行研究。方法先合成rutaecarpine,再进行N-取代反应,合成目标化合物,并测试所合成化合物的体外抗血小板聚集活性。结果与结论共合成6个未见文献报道的新化合物,它们的结构经1H-NMR和MS确证。初步体外药理实验结果显示:有4个新化合物对血小板聚集的抑制作用强于母体化合物rutaecarpine。     

Immunosuppressive effects of rutaecarpine in female BALB/c mice

Rutaecarpine is a major quinazolinocarboline alkaloid isolated from Evodia rutaecarpa. It was reported to possess a wide spectrum of pharmacological activities, such as vasodilation, antithrombosis, and anti-inflammation. In the present study, adverse effects of rutaecarpine on immune functions were determined in female BALB/c mice. Rutaecarpine had no effects on hepatotoxicity parameters in mice, as measured by serum activities of aminotransferases. Meanwhile, rutaecarpine significantly decreased the number of antibody-forming cells and caused weight decrease in spleen in a dose-dependent manner, when mice were administered with rutaecarpine at 10mg/kg, 20mg/kg, 40 mg/kg or 80 cmg/kg once intravenously. In addition, rutaecarpine administered mice exhibited reduced splenic cellularity, decreased numbers of total T cells, CD4(+) cells, CD8(+) cells, and B cells in spleen. IL-2, interferon-gamma and IL-10 mRNA expressions were suppressed significantly by rutaecarpine treatment. The number of CD4(+)IL-2(+) cells was reduced significantly following administration of mice with rutaecarpine. Furthermore, rutaecarpine caused the cell cycle arrest in G(0)+G(1) phase in a dose-dependent manner. Rutaecarpine caused significant inductions of hepatic cytochrome P450 (CYP) 1A, 2B, and 2E1 activities dose-dependently. In the splenic lymphocyte proliferation assay, rutaecarpine inhibited proliferation by LPS and Con A ex vivo, whereas it had no effects on in vitro proliferation. These results suggested that a single bolus intravenous injection of rutaecarpine from 20mg/kg might cause immunosuppressive effects, and that rutaecarpine-induced immunosuppression might be mediated, at least in part, through the inhibition of cytokine production and cell cycle arrest in G(0)+G(1) phase, and caused possibly by mechanisms associated with metabolic activation.     

Alteration of the pharmacokinetics of theophylline by rutaecarpine, an alkaloid of the medicinal herb Evodia rutaecarpa, in rats

Rutaecarpine is a main active alkaloid present in the medicinal herb, Evodia rutaecarpa. The cytochrome P450 (CYP) 1A2 substrate, theophylline, is an important therapeutic agent for the treatment of asthma, but has a narrow therapeutic index. To evaluate the pharmacokinetic interaction of theophylline with rutaecarpine, the effects of rutaecarpine on CYP1A2 activity and theophylline pharmacokinetics were investigated. Oral treatment of Sprague-Dawley rats with 50 mg kg(-1) rutaecarpine for three days through a gastrogavage caused a 4- and 3-fold increase in liver microsomal 7-ethoxyresorufin O-deethylation (EROD) and 7-methoxyresorufin O-demethylation activity, respectively. In the kidney, rutaecarpine treatment caused a 3-fold increase in EROD activity. In the lungs, EROD activity was elevated from an undetectable to a detectable level by rutaecarpine. Pharmacokinetic parameters of theophylline were determined using a microdialysis sampling method. Rutaecarpine pre-treatment increased the clearance of theophylline in a dose-dependent manner. Pre-treatment of rats with 50 mg kg(-1) rutaecarpine caused a 3-fold increase in theophylline clearance and a 70%, 68% and 68% decrease in the area under the concentration-time curve (AUC), mean residence time (MRT) and half-life, respectively. These results demonstrated that rutaecarpine treatment elevated CYP1A2 catalytic activity and theophylline excretion in rats. In patients taking theophylline, adverse effects might be noticed when a rutaecarpine-containing herbal preparation is used concomitantly.     

Rutaecarpine attenuates osteoclastogenesis by impairing macrophage colony stimulating factor and receptor activator of nuclear factor κ‐B ligand‐stimulated signalling pathways

Rutaecarpine is a major alkaloid isolated from Evodia rutaecarpa. Here, we investigated the effects of rutaecarpine on osteoclast differentiation induced by macrophage colony stimulating factor (M‐CSF) and receptor activator of nuclear factor κ‐B ligand (RANKL) in bone marrow‐derived macrophages (BMMs). Treatment with rutaecarpine significantly inhibited osteoclastogenesis and prevented bone resorption of BMM‐derived osteoclasts. Mechanistically, rutaecarpine decreased the protein level of nuclear factor of activated T cells cytoplasmic‐1 (NFATc1) and the phosphorylation of other signalling pathways during the osteoclast differentiation. Thus, rutaecarpine may be useful as a therapeutic agent for the treatment of bone diseases.