延胡索提取物在小鼠体内的药代动力学研究

发布时间：2018-06-24 09:56

本文选题：延胡索 + 生物碱　；参考：《浙江大学》2017年硕士论文

【摘要】：延胡索具有悠久的药用历史,具有活血,行气,止痛的功能。生物碱是其主要的活性成分,除镇痛外,还具有抗抑郁、抗肿瘤、抗菌、抗炎、抗氧化等多种药理作用。许多含延胡索的制剂在临床上已被用于治疗头痛、冠心病、胃肠炎、胃溃疡等多种疾病,对其活性成分在体内的血药浓度和组织分布测定有利于药物的合理使用。本研究建立了快速、灵敏、可同时定量14种生物碱—1个阿朴啡类：氧化海罂栗碱(OXO),1个原阿片碱类：原阿片碱(PRO),5个叔胺碱类：延胡索甲素(COR)、延胡索乙素(THP)、四氢小檗碱(4H-BER),四氢非洲防己胺(4H-COL)、四氢黄连碱(4H-COP)和7个季铵碱类：非洲防己碱(COL)、巴马汀(PAL)、小檗碱(BER)、表小檗碱(EPI)、黄连碱(COP)、药根碱(JAR)、去氢延胡索甲素(DH-COR)的生物样品HPLC-MS/MS分析方法,并应用该方法对经口服给予延胡索提取物后14种生物碱在小鼠血浆、脑、心脏、肝脏和肾脏药代动力学特征进行了研究,主要结果如下：1.液质联用法测定小鼠血浆中14种生物碱的方法建立与药代动力学研究本研究建立并验证了快速、灵敏、可同时定量14种生物碱在小鼠血浆中的HPLC-MS/MS分析方法。色谱柱为ZORBAX Eclipse XDB-C18 (2.1 × 100 mm,3.5 μm),流动相为乙腈(0.1%甲酸)—水(0.1%甲酸),流速为0.3 mL/min进行梯度洗脱。以氯化两面针碱作为内标,采用MRM模式检测。按照中国药典四部中的生物样品定量分析方法验证指导原则(2015年版)和美国FDA生物分析方法验证指导原则(2013年版)进行方法学验证,符合限度规定。小鼠灌胃给予1.25 g/kg延胡索提取物后于0～24 h内的不同时间点采集血浆样本,并在本实验条件下检测,BER和EPI低于定量下限。通过比较12种生物碱的主要药代动力学参数发现,延胡索提取物中的大多数生物碱能快速被吸收入血,THP在血中消除最快。2.液质联用法测定小鼠4种组织中14种生物碱的方法建立与组织分布研究本研究在小鼠血浆分析方法的基础上,建立和验证了快速、灵敏、可同时定量小鼠组织中14种生物碱的HPLC-MS/MS分析方法,样品在不同的储存条件下保持稳定,验证结果均在可接受水平。该分析方法成功应用于小鼠灌胃1.25 g/kg延胡索提取物后脑、心脏、肝脏、肾脏组织中14种生物碱的分布,并得出组织中药代动力学参数。结果表明延胡索提取物中的大多数生物碱能迅速分布到4种组织中,所有生物碱在肝脏中的分布量最大。THP、COP和PRO在脑组织中的分布量较高；COP、DH-COR和JAR在心脏组织中的分布量较高；COP、DH-COR和PRO在肝脏中分布量较高,迅速分布于肝脏的叔胺碱浓度逐渐降低至LLOQ,未出现明显的再次分布或转化生成,而季铵碱类生物碱较缓慢的分布于肝脏,并很可能存在再分布或药物转化；COP、DH-COR和PRO在肾脏中分布量较高。
[Abstract]:Corydalis corydalis has a long medicinal history, has the function of promoting blood circulation, Qi, pain relief. Alkaloids are the main active components of alkaloids. Besides analgesia, alkaloids also have many pharmacological effects, such as antidepressant, antitumor, antimicrobial, anti-inflammatory, anti-oxidation and so on. Many preparations containing Corydalis corydalis have been used to treat headache, coronary heart disease, gastroenteritis, gastric ulcer and other diseases. The determination of blood drug concentration and tissue distribution of its active components is beneficial to the rational use of drugs. In this study, a rapid and sensitive method was established. Simultaneously quantifiable 14 alkaloids-1 aporphine: OXO, 1 proopiate: pro, 5 tertiary amine alkaloids: Corydalis A (cor), tetrahydropalmatine (THP), tetrahydroberberine (4H-BER), tetrahydroAfrican prohexamine (4H-COL), tetrahydroprotophan (4H-COL), tetrahydropalmatine (THP), tetrahydropalmatine (4H-COL) HPLC-MS-MS analysis of hydrocortisine (4H-cop) and seven quaternary ammonium bases: African Tetrandrine (Col), berberine (pal), berberine (BER), epiberberine (EPI), Coptis base (cop), root alkaloid (jar), dehydrocortisone (DH-COR). The pharmacokinetic characteristics of 14 alkaloids in plasma, brain, heart, liver and kidney of mice after oral administration of Corydalis extract were studied. The main results are as follows: 1. Establishment and Pharmacokinetics of 14 Alkaloids in Mouse Plasma by liquid / Mass Spectrometry the method of HPLC-MS / MS for the simultaneous determination of 14 alkaloids in mouse plasma was established and verified. The chromatographic column was ZORBAX Eclipse XDB-C18 (2. 1 脳 100mm) 3.5 渭 m), mobile phase consisted of acetonitrile (0.1% formic acid) -water (0.1% formic acid), and the flow rate was 0.3 mL / min for gradient elution. The internal standard of nitidine chloride was detected by MRM mode. In accordance with the guidelines for the verification of quantitative analysis of biological samples in the four parts of the Chinese Pharmacopoeia (2015 edition) and the FDA guidelines for the Verification of Biological Analysis methods (2013 edition), the methodology verification is carried out in accordance with the limits stipulated. Plasma samples were collected at different time points within 24 hours after administration of 1.25 g/kg extract of Corydalis corydalis by intragastric perfusion in mice, and the levels of BER and EPI were lower than the lower limit of quantification under this experimental condition. By comparing the main pharmacokinetic parameters of 12 alkaloids, it was found that most of the alkaloids in the extract of Corydalis corydalis could be quickly absorbed into blood to eliminate THP. Establishment and tissue Distribution of 14 Alkaloids in four kinds of tissues of mice; based on the method of plasma analysis in mice, a rapid and sensitive method was established and verified. HPLC-MS / MS method for simultaneous quantification of 14 alkaloids in mouse tissues showed that the samples remained stable under different storage conditions and the results were at acceptable level. The method was successfully applied to the distribution of 14 alkaloids in the brain, heart, liver and kidney of mice fed with 1.25 g/kg extract of Corydalis corydalis, and the pharmacokinetic parameters of traditional Chinese medicine were obtained. The results showed that most of the alkaloids in the extract of Corydalis corydalis could be distributed rapidly in four kinds of tissues. The distribution of all alkaloids in the liver was the highest. The distribution of cop and pro in brain tissue was higher than that of COPDH-COR and jar in heart tissue. The concentration of tertiary amine alkaloids rapidly distributed in the liver decreased to LLOQ, and there was no obvious redistribution or transformation, while the alkaloids of quaternary ammonium alkaloids distributed slowly in the liver. It is very likely that there is a higher distribution of COPH-COR and pro in the kidney.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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