【作者】 孙晓博；

【导师】 金描真；

【作者基本信息】 广东药学院 ， 药剂学， 2009， 硕士

【摘要】 血竭（Sanguis Draconis）为棕榈科植物麒麟竭（Deamonorops draco BL）及同属其他种植物和龙舌兰科植物剑叶龙血树（Dracaena Cochinchinensis（Lour.）S.C.Chen）及同属植物的果实或含树脂木材的乙醇提取物,植物来源分属4科5属的20余种植物,作为传统中药使用已有1500多年的历史。至1971年开发成功了国产血竭,为传统中药血竭的国产化奠定了基础。国产血竭与进口血竭化学成分不同,但都具有活血化瘀、消肿止痛、收敛止血的功效,尤其是近年来采用国产血竭治疗心脑血管系统疾病取得了较好的疗效。本论文以国产广西龙血竭为研究对象,以龙血素A为指标性成份,对龙血竭的在体内的药物动力学情况进行了研究。目前,国内外已有部分关于龙血素A含量测定方法的报道,本实验建立了龙血素A在血浆中及在透析液中的含量分析方法,结合平衡透析法考察了龙血素A的人血浆蛋白结合率。采用高效液相色谱法,测定大鼠、比格犬在灌胃给药后,全血中龙血素A的含量。同时,采用该方法测定了大鼠灌胃给药后各脏器组织中不同时间点龙血素A的含量。本课题为血竭的临床合理用药提供了依据。全文由以下四部分组成:（一）绪论概述血竭的生药学、化学成分、药理作用及其药物动学的研究现状。（二）龙血素A的血浆蛋白结合率建立了龙血素A在血浆中的测定方法,采用平衡透析法对龙血素A人血浆蛋白结合率进行测定。结果低、中、高三种浓度下,龙血素A的血浆蛋白结合率分别为79.5±3.1%,80.7±1.1%,80.3±0.9%,说明龙血素A与血浆蛋白的结合能力较强。本文建立的方法灵敏度高,重现性好,操作简单,能满足分析要求。（三）龙血素A在大鼠体内的药物动力学研究用HPLC法测定大鼠全血中龙血素A的含量,对其在大鼠体内的药物动力学进行研究。采用Diamonsil ODS C18色谱柱,以乙腈:1%冰醋酸（35:65）为流动相,流速1.2ml·min^-1,检测波长280nm,柱温25℃。按每100g大鼠给予龙血竭250mg,灌胃给药,测定不同时间点的血药浓度,并以DAS软件计算其药动学参数。龙血素A检测限（LOD）为0.025μg·mL^-1;在0.051～4.836μg·mL^-1范围内线性良好,r为0.9983;低、中、高浓度的绝对回收率及方法回收率均在80%以上。日内及日间RSD均小于8%。龙血素A的Ka为2.09h^-1 ;t_1/2（Ka）为0.33h;Ke为0.32 h^-1 ;t_1/2（Ke）为2.19h;tmax为2.06h;Cmax为0.67μg·mL^-1;AUC为2.08μg·mL^-1·h。龙血素A在大鼠体内符合线性动力学一室开放模型,吸收较快,消除也较快。研究龙血竭中龙血素A在大鼠体内的组织分布。按每100g大鼠给予龙血竭250mg,灌胃给药,用HPLC法测定不同时间点组织中龙血素A的浓度。结果发现龙血素A在肝脏及肾脏浓度最高,其他脏器依次是肺、脾、心、脑。证明龙血素A在大鼠体内主要分布于血流量大的脏器组织,同时也说明龙血素A可以透过血脑屏障。（四）龙血素A在比格犬体内的药物动力学研究用HPLC法测定比格犬全血中龙血素A的含量,并对其在比格犬体内的药物动力学情况进行研究。采用Diamonsil ODS C₁₈色谱柱,以乙腈:1%冰醋酸（35:65）为流动相,流速1.2ml·min^-1,检测波长280nm,柱温25℃。按1kg比格犬给予龙血竭提取物250mg,灌胃给药,测定不同时间点的血药浓度,并以DAS软件计算其药动学参数。龙血素A检测限（LOD）为0.012μg·mL^-1;在0.025～2.05μg·mL^-1范围内线性良好,r为0.9984;低、中、高浓度的绝对回收率及方法回收率均在80%以上。日内及日间RSD均小于8%。龙血素A的Ka为0.47h^-1 ;t_1/2（Ka）为1.49h;Ke为0.37 h^-1 ;t_1/2（Ke）为1.86h;tmax为3.00h;Cmax为0.18μg·mL^-1,AUC为1.07μg·mL^-1.h。更多还原

【Abstract】 Sanguis Draconis is mainly made of the ethanol extractive of resin or fruits from deamonorops draco BL. and Dracaena Cochinchinensis （Lour.）S. C. Chen, originated from more than 20 kinds of plants which belong to 4 branches 5 categories. It has more than 1500 years in its application as precious traditional herb. Sanguis Draconis made in China developed from 1971. It establishes the foundation of nationalization of traditional Chinese herb, Sanguis Draconis. Domestic and import Sanguis Draconis distinguish in chemical constituents, but their medical activities are similar, including quickening the blood and transforming stasis, detumenscen and relieving pain, constringency and stanch bleeding. At present, Sanguis Draconis made in China makes better healing efficacy in angiocardiopathy.Study on pharmacokinetics of Loureirin A f Sanguis Draconis. The metabolism of Loureirin A in rats after oral administration,with the parameters of pharmacokinetics calculated with DAS pharmacokinetics program. After ig. Loureirin A in rats, the concentration in tissues was determined by HPLC.The main contents can be summarized as follows:1 IntroductionSummarized advances in studies on pharmacognosy, chemical constituents, pharmacological action and pharmacokinetics of Loureirin A of Sanguis Draconis.2 Determination of blood protein binding rate of Loureirin AThe blood protein binding rate of Loureirin A of rats and human were determined. The equilibrium dialysis combined with HPLC to determine the blood concentration and blood protein binding rate of Loureirin A was carried out. The blood protein binding rates of Loureirin A at low, middle and high concentration were 79.5%, 80.7% and 80.3% respectively.3 Research on the pharmacokinetics of Loureirin A in rats To establish a RP—HPLC method for the determination of Loureirin A in rat blood for pharmacokinetics study after a single dose of Dracaena cochinehinesis（Lour.）S.C.Chen in rats. The samples were mixed with acetonitrile and ethyl acetate. The organic layer separated was blown to dry and dissolved reconstituted in methanol.The HPLC separation was perform ed on a Cl8 column and the UV detector wavelength was set at 280 nm.The blood samples were analyzed after an oral administration of Dracaena cochinehinesis（Lour.）S.C.Chen to rats (2.5g·kg^-1 ),the pharmacokinetic parameters were valuated by DAS software.Good linearities of Loureirin A was obtained over the range of 0.051～4.836μg·mL^-1 in blood.The recoveries of Loureirin A was over 80%.The precisions of intra-day and inter—day were all less than 8%.The main pharmacokinetics parameters were as follows:Ka of Loureirin A was 2.09h^-1; t_1/2（Ka） 0.33h and Ke was 0.32h-1 t_1/2（Ke） 2.19h;tmax was 2.06h;Cmax 0.64μg·mL^-1 and AUC 2.08μg·mL^-1·h. The method is sensitive,accurate and reproducible,and can be applied to the Loureirin A in blood.The sharmacokinetics characteristics showed Loureirin A was absorbed rapidly and eliminated soon.Study on the tissue distribution of Loureirin A in rats. After ig. administration of Loureirin A at does of 250mg/100g in rats, the tissue concentrations at different time points were determined by HPLC.After ig. administration, the main tissue distribution of Loureirin A concentrations in the liver and kidney were the highest and those in the spleen,heart,lung,brain,stomach,fat and muscle decreased sequentially.4 Research on the pharmacokinetics of Loureirin A in dogs To establish a RP—HPLC method for the determination of Loureirin A in dogs’blood for pharmacokinetics study after a single dose of Dracaena cochinehinesis（Lour.）S.C.Chen . The samples were mixed with acetonitrile and ethyl acetate. The organic layer separated was blown to dry and dissolved reconstituted in methanol.The HPLC separation was performed on a C₁₈ column and the UV detector wavelength was set at 280 nm. The blood samples were analyzed after an oral administration of Dracaena cochinehinesis（Lour.）S.C.Chen to dogs (0.25g·kg^-1 ),the pharmacokinetic parameters were valuated by DAS software.Good linearities of Loureirin A was obtained over the range of Good linearities of Loureirin A was obtained over the range of 0.025～2.05μg·mL^-1 in blood.The recoveries of Loureirin A was over 80%.The precisions of intra-day and inter—day were all less than 10%.The main pharmacokinetics parameters were as follows Ka of Loureirin A was0.47h^-1; t_1/2（Ka） was 1.49h and t_1/2（Ke） was 1.86h; Ke was 0.37 h^-1;tmax was 3.00h;Cmax 0.18μg·mL^-1 and AUC 1.07μg·mL^-1h. CONCLUSION The method is sensitive,accurate and reproducible,and can be applied to the Loureirin A in blood.The sharmacokinetics characteristics showed Loureirin A was absorbed and eliminated slowly in dogs.更多还原