反相高效液相色谱法测定骆驼蓬总碱片中去氢骆驼蓬碱和骆驼蓬碱的含量

目的 :建立RP HPLC法测定骆驼蓬总生物碱片中去氢骆驼蓬碱和骆驼蓬碱含量的方法。方法 :以甲醇 0 .0 1mol·L-1硫酸铵溶液 二乙铵 (6 0∶40∶0 .4,用磷酸调 pH至 (3.8± 0 .1)为流动相 ,进样量 2 0 μl,分别在 32 0 ,372nm处用RP HPLC法测定去氢骆驼蓬碱和骆驼蓬碱的含量。结果 :去氢骆驼蓬碱和骆驼蓬碱分别在浓度为 2～ 2 5 μg·ml-1的范围内与峰面积成良好的线性关系。方法的回收率分别为 (10 1.8± 1.8) %和 (10 1.3± 1.4) %。结论 :该法简便 ,准确 ,可靠     

高效液相色谱法测定 哈林胶囊中去氢骆驼蓬碱与骆驼蓬碱含量

［摘要］目的建立高效液相色谱（HPLC）法测定哈林胶囊中去氢骆驼蓬碱和骆驼蓬碱含量的方法。方法以甲醇 0.01moL·L 1硫酸铵溶液（40：60，用磷酸调pH至4.0）为流动相，进样量10 μL，在320 nm处用HPLC法测定去氢骆驼蓬碱和骆驼蓬碱的含量。结果去氢骆驼蓬碱和骆驼蓬碱浓度均在1～50 μL·mL 1范围内与峰面积成良好的线性关系。回收率分别为 98.37% 和 99.28%，RSD分别为1.39%和0.82%。结论该方法简便，准确，可靠。     

去氢骆驼蓬碱衍生物的合成和抗肿瘤活性研究

目的 合成去氢骆驼蓬碱衍生物并研究其抗肿瘤活性。方法 以去氢骆驼蓬碱为原料,在NaH催化下先用卤代烷烃对9位氮原子进行烷基化,然后用溴化苄将2位氮原子进行季铵化,得到一系列去氢骆驼蓬碱衍生物,用 MTT 法考察其对肿瘤细胞的抑制作用。结果 合成了6个新的去氢骆驼蓬碱衍生物,结构经1H-NMR、MS和红外光谱确证。与去氢骆驼蓬碱相比,所合成的化合物均有明显的抗肿瘤活性。结论 初步药理实验结果表明去氢骆驼蓬碱9-苯丙基取代能够明显提高抗肿瘤效果。     

去氢骆驼蓬碱脂质体的研究

用正文试验法筛选出去氢骆驼蓬碱（ｈａｒｍｉｎｅ，１）脂质体的处方组成，并采用醚滴注－超声法制备脂质体。其包封率为２９．４２±０．７４％，粒径为０．１０±０．０３μｍ。与水溶液比较，它能显著提高１在小鼠肺、脾、肾和肝等脏器中的分布总量。给家兔ｉｖ和ｉｇ上述两种剂型后，１在兔体内的药时过程均符合二室模型。     

去氢骆驼蓬碱聚合物胶束的制备工艺优化及体外释放的研究

目的：研究星点设计-响应面法优化去氢骆驼蓬碱N-癸酰-N-三甲基壳聚糖胶束（harmine loaded N-Decanoate-N-trimethyl chitosan micelles,HM-De-TMC-MIC）的处方优化,并考察在不同介质中HM-De-TMC-MIC的体外释放。方法：以薄膜分散法制备HM-De-TMC-MIC;以粒径、多分散系数、包封率和载药量为指标,通过单因素考察和星点设计-响应面法综合考察药物与载体质量比和复水体积对HM-De-TMC-MIC的影响,并遴选其最优处方。在不同pH的释放介质中,分别考察HM-De-TMC-MIC和HM的体外释放。结果：筛优处方药物与载体质量比为3.6∶10,复溶水体积为6 mL;以最优处方制备的HM-De-TMC-MIC粒径为（148.2±5.0）nm,多分散系数为0.198±0.045,包封率（89.80±0.19）%,载药量（22.79±0.05）%,形态圆整。体外释放试验结果表明,HM-De-TMC-MIC的释放曲线遵循Higuchi方程,与HM溶液相比其释放较为缓慢,并呈现pH敏感释药行为。结论：以星点设计-响应面法优化的HM-De-TMC-MIC具有较好包封率和载药量,粒径分布均匀,具有明显的缓释性。     

去氢骆驼蓬碱脂质体的制备和体外释放特性

目的：研究去氢骆驼蓬碱（harmine,HM）脂质体的制备工艺和体外释放特性。方法：运用薄膜分散-pH值梯度法制备HM．脂质体以及高速离心法分离脂质体与游离药物,并测定其包封率;借助综合评分法,评价其粒径、多分散系数、包封率、载药量指标;运用正交优化实验法考察磷脂-胆固醇与药-脂比、超声时间、外相pH值对脂质体的影响,述选最优工艺处方,评价脂质体与原料药的体外释放情况。结果：最优处方因素为磷脂-胆固醇比值为4：1,超声时间为300S,药-脂比值为1：5,外相pH值为6,8,即X13X23X32X43,经实验验证其粒径为（155.0±14．5）nm,多分散系数为（0．148±0．011）,包封率为（80．90±0．01）％,载药量为（11．16±0．01）％;其原料药0．5h累积释放百分比大于50％,不到2h已全部释放,而优化后的脂质体在1h内其累积释放百分比大于50％,4h后释放完成。结论：采用薄膜分散-pH值梯度法,以最优处方制得HM-脂质体,其粒径大小适中、形态均匀,包封率和载药量相对较高,体外释放显示具有较好的缓释特性。     

骆驼蓬属植物种子中去氢骆驼蓬碱和骆驼蓬碱的HPLC-荧光检测法测定

建立了HPLC-荧光检测法测定不同产地骆驼蓬、骆驼蒿、多裂骆驼蓬种子（共15个样品）中去氢骆驼蓬碱（1）和骆驼蓬碱（2）的含量。采用C18色谱柱，以乙腈-醋酸铵缓冲液（19：81）为流动相，激发波长为250nm、发射波长为460nm。结果显示，骆驼蓬中1的含量（3．6％～5．0％）高于2（2．8％～3．5％）；多裂骆驼蓬则相反，1、2的含量分别为1．6％～2．0％和2．3％～3．1％；骆驼蒿中2的含量较高（4．4％），但1含量低于检测限。     

Streptomyces griseus ATCC 13273对去氢骆驼蓬碱的转化研究

利用微生物转化的方法,选取灰色链霉菌(Streptomyces griseus ATCC 13273)为转化菌株,对去氢骆驼蓬碱的转化进行研究。该菌株可以将去氢骆驼蓬碱转化为两种转化产物(记为H1和H2),通过MS、1H-NMR和13C-NMR分析,转化产物分别鉴定为哈尔酚和N-羟基去氢骆驼蓬碱。并对转化条件进行初步优化,优化后的转化条件为:初始pH 8.0,两步活化法以4%的接种量转接,48 h后加入底物(以N,N-二甲基甲酰胺溶解),发酵液中去氢骆驼蓬碱终浓度为0.25 mmoL/L,发酵时间为5 d。转化条件优化后H1和H2的产率分别达到8.70%和12.76%;该研究丰富了S.griseus ATCC 13273的作用底物范围,为哈尔酚和N-羟基去氢骆驼蓬碱的合成提供了一条新途径。     

γ-去氢骆驼蓬碱等咔啉类生物碱的辐射防护作用

γ- Harmine(Ⅰ),’harmine(Ⅱ )and harmaline(Ⅲ )were isolated from PeganumHarmala L.( Zygophylaceae).Tests were conducted with mice to detect whether γ- harmine( a newcompound), harmine,harmof(Ⅳ)and harmalol(Ⅴ) are effective radioprotective compounds againstγ-ray irradiation, Intraperitoneal injection of the hydrochlorides of the four alkaloids 50～80mg·kg^-1×1 in NIH male mice 30～45 minutes before 8.6～9. 7 Gy whole body ⁶⁰Co irradiationsignificantly increased the survival effects(1. 33～2. 61)and 30-day survivai rate in comparison withcontrol mice.The results indicatethat γ- harmine exhibited relatively good radioprotective effect.γ-harmine is the first alkaloid isolated from a plant having ptotective effects against whole-body lethal irradiation in mice.     

γ－去氢骆驼蓬碱等咔啉类生物碱的辐射防护作用

γ－去氢骆驼蓬碱等咔啉类生物碱的辐射防护作用利国威，桑培根，潘国英（中山医科大学肿瘤防治中心广州５１００６０）从新疆产蒺藜科植物骆驼蓬（ＰｅｈａｎｕｍｈａｒｍａｌａＬ．）的种子中分离得到３个化合物，其中γ－去氢骆驼蓬碱（γ－ｈａｒｍｉｎｅ，Ⅰ）为新化...     

Exploring the mechanism of Peganum harmala L. seeds on hepatocellular carcinoma based on network pharmacology and molecular docking

Peganum harmala L. is a medicinal plant, and its seeds have been used to treat gastrointestinal cancer and malaria for a long time in North-Western China. In the present study, we aimed to probe the potential molecular targets and pharmacological mechanisms of Peganum harmala L. seeds (PHS) on hepatocellular carcinoma (HCC) using network analysis and molecular docking. First, the chemical ingredients of PHS were obtained from TCMID and BATMAN-TCM databases, and the effective ingredients were screened by SwissADME. Furthermore, the target information of the effective ingredients was acquired from PharmMapper and SwissTargetPrediction databases. Secondly, HCC-related targets were obtained from Liverome, DisGeNET, and GeneCards databases. The intersection with the PHS was obtained. The "compounds-targets" was drawn using Cytoscape software, and PPI network diagrams were drawn using their intersection targets. GO analysis and KEGG enrichment analysis were carried out using the DAVID database. Finally, molecular docking was conducted between protein receptors and the active components using AutoDockTools. Our results showed 105 intersection targets of PHS with HCC. Moreover, 10 core targets included ALB, AKT1, EGFR, CASP3, SRC, ESR1, MAPK3, MMP9, ANXA5, and MAPK1. Besides, 404 GO functional annotations were obtained, including 287 biological processes, 37 cell compositions, and 80 molecular functions. In addition, 110 signaling molecules and pathways, including chemical oncogene receptors, PI3K-Akt pathway, HCC, and hepatitis B, were identified. The molecular docking results showed that the binding energies of the 10 core targets and the 12 active components were all less than –5 kcal/mol. In conclusion, this study expounded on the "component-target-pathway" interaction mechanism of PHS for the treatment of HCC, and it also provided a scientific basis for the clinical application of PHS.     

Vasorelaxant effects of harmine and harmaline extracted from Peganum harmala L. seeds in isolated rat aorta.

The present work describes the mechanisms involved in the vasorelaxant effect of harmine and harmaline. These alkaloids induce in a dose-dependent manner the relaxation in the aorta precontracted with noradrenaline or KCl. However, the removal of endothelium or pre-treatment of intact aortic ring with L-NAME (inhibitor of NOSe synthetase) or with indomethacin (non-specific inhibitor of cyclo-oxygenase), reduces significantly the vasorelaxant response of harmaline but not harmine. According to their IC50 values, prazosin (inhibitor of alpha-adrenorecepteors) reduces the vasorelaxant effect only of harmaline, whereas, pre-treatment with IBMX (non-specific inhibitor of phosphodiesterase) affects both the harmaline and harmine-responses. Inhibitions of L-type voltage-dependent Ca2+ channels (VOCs) in endothelium-intact aortic rings with diltiazem depress the relaxation evoked by harmaline as well as by harmine. Pre-treatment with harmaline or harmine (3, 10 or 30 microM) shifted the phenylephrine-induced dose response curves to the right and the maximum response was attenuated indicating that the antagonist effect of both alkaloids on alpha1-adrenorecepteors was non-competitive. These two alkaloids also exert an antioxidant activity by scavenging the free radical generated by DPPH. Therefore, the present results suggest that the vasorelaxant effect of harmaline but not harmine is related to its action on the prostacyclin pathway and on the endothelial cells to release NO. However, both alkaloids can act as blockers VOCs, as inhibitors of phosphodiesterase resulting in an increase of the second messenger (cAMP and cGMP) levels and finally reduce the levels of free radicals in tissues.     

HPLC method for the analysis of harmol,harmalol, harmine and harmaline in the seeds of Peganum harmala L

A simple and sensitive method for separation and determination of harmol, harmalol, harmine and harmaline has been developed and validated. Harmol, harmalol, harmine and harmaline were separated using a Metasil ODS column by isocratic elution with flow rate 1.5 ml/min. The mobile phase composition was Isopropyl alcohol-Acetonitrile-Water-Formic acid (100:100:300:0.3) (v/v/v/v) and pH adjusted 8.6 with triethylamine. Spectrophotometric detection was carried out at 330 nm. The linear range of detection for harmol, harmalol, harmine and harmaline were between 9.375-250, 30.750-246, 31.250-500 and 31.000-248 microg/ml, respectively. The method described was suitable for the determination of harmol, harmalol, harmine and harmaline in the seeds of Peganum harmala L.     

Protective effects of Peganum harmala L. extract, harmine and harmaline against human low-density lipoprotein oxidation

Oxidative modification of low-density lipoprotein (LDL) particles has been implicated in the process of atherogenesis. Antioxidants that prevent LDL from oxidation may reduce atherosclerosis. We have investigated the protective effect of Peganum harmala-extract (P-extract) and the two major alkaloids (harmine and harmaline) from the seeds of P. harmala against CuSO4-induced LDL oxidation. Through determination of the formation of malondialdehyde (MDA) and conjugated diene as well as the lag phase, the extract (P-extract) and compounds were found to possess an inhibitory effect. Moreover, harmaline and harmine reduced the rate of vitamin E disappearance and exhibited a significant free radical scavenging capacity (DPPH*). However, harmaline had a markedly higher antioxidant capacity than harmine in scavenging or preventive capacity against free radicals as well as inhibiting the aggregation of the LDL protein moiety (apolipoprotein B) induced by oxidation. The results suggested that P. harmala compounds could be a major source of compounds that inhibit LDL oxidative modification induced by copper.     

A rapid and simple method for the determination of psychoactive alkaloids by CE‐UV: application to Peganum Harmala seed infusions

The β‐carboline alkaloids of the harmala (HAlks) group are compounds widely spread in many natural sources, but found at relatively high levels in some specific plants like Peganum harmala (Syrian rue ) or Banisteriopsis caapi . HAlks are a reversible Mono Amino Oxidase type A Inhibitor (MAOI) and, as a consequence, these plants or their extracts can be used to produce psychotropic effects when are combined with psychotropic drugs based on amino groups. Since the occurrence and the levels of the HAlks in natural sources are subject to significant variability, more widespread use is not clinical but recreational or ritual, for example B . caapi is a known part of the Ayahuasca ritual mixture. The lack of simple methods to control the variable levels of these compounds in natural sources restricts the possibilities to dose in strict quantities and, as a consequence, limits its use with pharmacological or clinical purposes. In this work, we present a fast, simple, and robust method of quantifying simultaneously the six HAlks more frequently found in plants, i.e., harmine, harmaline, harmol, harmalol, harmane, and norharmane, by capillary electrophoresis instruments equipped with the more common detector UV. The method is applied to analyze these HAlks in P . Harmala seeds infusion which is a frequent intake form for these HAlks. The method is validated in three different instruments in order to evaluate the transferability and to compare the performances between them. In this case, harmaline, harmine, and harmol were found in the infusion samples. Copyright © 2016 John Wiley & Sons, Ltd.     

不同炮制方法对骆驼蓬子总生物碱浸出率的影响

Total alkaloids extracted from Peganum Harmala proccsscd dlfferently was determined by the first derivative spetrometry Results from the variance analysis showed that the effect of the dlffercnt processing ways on extraction of total alkaloids from seed     

黄荆子亲脂性化学成分研究

目的研究马鞭草科牡荆属植物黄荆果实亲脂性化学成分。方法采用硅胶柱层析法、ODS柱层析法、Sephadex LH-20柱层析等分离方法对黄荆子亲脂性化学成分进行分离纯化,根据理化性质和波谱数据进行结构鉴定。结果分离得到7个单体化合物,分别为abietatrien-3β-ol(Ⅰ)、vitexifolin C(Ⅱ)、phytol(Ⅲ)、obtusalin(Ⅳ)、L-芝麻脂素(Ⅴ)、邻苯二甲酸二丁酯(Ⅵ)、β-谷甾醇(Ⅶ)。结论化合物为Ⅰ,Ⅱ,Ⅲ首次从该植物中分离得到。     

骆驼蓬种子提取物及其β咔保啉生物碱对DNA拓扑异构酶Ⅱ活性的抑制作用

目的探讨去氢骆驼蓬碱、骆驼蓬碱、骆驼蓬总碱及哈尔满碱(止咳药用植物)对DNA拓扑异构酶Ⅱ活性的抑制作用。方法从体外培养的Q3肝癌细胞中,提取分离DNA拓扑异构酶Ⅱ;以阿霉素为阳性对照,用琼脂糖凝胶电泳法检测药物对DNA拓扑异构酶Ⅱ的作用。结果骆驼蓬总碱、去氢骆驼蓬碱、骆驼蓬碱及哈尔满碱对DNA拓扑异构酶Ⅱ活性均有一定的抑制作用。结论对DNA拓扑异构酶Ⅱ活性的抑制作用是骆驼蓬总碱、去氢骆驼蓬碱、骆驼蓬碱等生物碱抗癌作用和细胞毒作用的机制之一。     

Interspecies metabolic diversity of harmaline and harmine in in vitro 11 mammalian liver microsomes

The β‐carboline alkaloids harmaline and harmine are widely present in hallucinogenic plants with great potential for treating depression, Parkinson's disease, and Alzheimer's disease. The present study was to elucidate metabolic difference of harmaline and harmine in 11 mammalian liver microsomes in order to quantitate species‐specific metabolic profiles. Using the probe substrate reaction, the enzymatic activities for 8 CYP450 isozymes of 11 liver microsomes were characterized. Combining ultra performance liquid chromatography combined with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry (UPLC‐ESI‐Q/TOF‐MS) and ultra performance liquid chromatography combined with electrospray ionization quadrupole tandem mass spectrometry (UPLC‐ESI‐MS/MS) methods, 18 metabolites for harmaline and 11 for harmine were identified. The metabolism patterns differences of them presented discrepancy in the quality and quantity of metabolites. It was notable that O ‐sulfate conjugation was detected in all species except sheep. The intrinsic clearance CL _{int , LM} values for the metabolites harmine and harmol in rabbits (37.5 and 42.4 μL/min/mg) were higher than those in other animals, while dogs (16.2 and 16.7 μL/min/mg) and humans (16.0 and 16.3 μL/min/mg) exhibited similar in vitro metabolic clearance. These observations suggested that harmaline and harmine were rapidly metabolized in liver microsomes of rat, mouse, and rabbit; moderately metabolized in human and dog; while weakly metabolized in sheep. Comprehensive analysis of the metabolism indicated that dogs and humans showed considerable similarity in the elimination of parent drugs, metabolic profiles, and catalytic processes. To summarize, these findings illustrated that in vitro studies of harmaline and harmine metabolic profiles in different species are helpful for the proper selection and interpretation of animal models for pharmacological and toxicological evaluation, and will ultimately provide useful guidance for the development of β‐carboline alkaloids. Copyright © 2016 John Wiley & Sons Ltd.