论文标题:莱格注射液的制备工艺及其药代动力学研究
Study on the Preparation Technology and Pharmacokinetics of the Intravenous Emulsion of Lig
论文作者 魏玉平
论文导师 任天池,论文学位 博士,论文专业 中药药剂学
论文单位 北京中医药大学,点击次数 97,论文页数 102页File Size1400k
2005-06-01论文网 http://www.lw23.com/lunwen_411899162/ 急性肾功能衰竭;莱格内酯;生物转化;药物代谢动力学;脂肪乳
Lig Lactone; intravenous emulsion ; acute renal failure(ARF) ; pharmacokinetic(PK);biological transformation
目的 莱格内酯是从植物中提取的活性成分,可对抗急性肾功能衰竭,目前国内外均无以治疗该疾病为目的的药物上市的报道。 方法 莱格内酯易于发生结构改变,一般的变化包括有①空气氧化;②热分解;③构型转换;④羟化及脱羟化等。鉴于不稳定,选择脂肪乳作为剂型,以制剂中莱格内酯的稳定性为评判指标,筛选出脂肪乳作为新制剂的剂型,并对制备工艺条件进行了考察,中试研究表明处方组成合理、工艺稳定、可控、易行。 建立莱格注射液的反相-高效液相色谱含量测定方法,并测定中试产品中莱格内酯的含量。系统研究了三批样品的物理和化学稳定性。 建立血中莱格内酯的含量测定方法。进行莱格注射液大、中、小三个剂量的大鼠体内药动学研究和一个剂量的重复给药的大鼠体内药动学研究。进行一个剂量的 Beagle犬体内药动学研究。并用 3p87 软件对采集的数据进行处理。 研究莱格内酯原形的粪、尿和胆汁的排泄情况。 研究大鼠的体外转化代谢情况,找出代谢途径。 研究莱格注射液对抗甘油造成的大鼠急性肾衰。 结果 (1)中试研究表明处方组成合理、工艺稳定、可控、易行。脂肪乳剂型解决了莱格内酯稳定性的问题,能够使药效成分在脂肪乳中保持稳定,制剂以市售包装在 6℃±2℃和 25℃±2℃保存一年各项指标均能达到要求,考虑到后者在放置一年时含量略有降低,确定莱格注射液应保存在 6℃±2℃的环境里。 (2)莱格注射液大、中、小三剂量于尾静脉注射后,在大鼠体内的药时行为符合三室模型(权重系数为 1/C2),莱格内酯到浅外室的分布半衰期 :t1/2pi约在 2～8min;莱格内酯到浅外室的分布半衰期:t1/2α约在 20～27min。消除半衰期t1/2β约在 330～619min。表观分布容积Vc为 0.23～0.62L/Kg,药物由中央室的消除速度常数K10约为1 .6*10~(-2)～5*10~(-2)(min-1),清除率CL(s)约为 5.21*10~(-3)～1.8*10~(-2),L/(kg*min),统计矩法计算结果显示大、中、小剂量的AUC值约为 2800、1695、756 (μg*min)/ml左右。 梯形法求得AUC值分别 2455、1538、695 (μg*min)/ml。 - 6 - 莱格注射液的制备工艺及其药代动力学研究(5) 大鼠分布实验显示莱格注射液注射后快速分布到体内器官组织,注射 10min后,其组织浓度分布由高向低的顺序如下: 血>胸腺>体脂>肠道>肾>子宫及卵巢>脑>胃>肺>前列腺>脾>肝>睾丸及附睾>骨骼肌。(6)莱格内酯的尿、粪和胆汁排泄研究结果表明,总量尚不足 10%。(7)莱格内酯具有很强的蛋白结合力,人血浆蛋白结合率在 97%以上。(8)以生理盐水加药、空白血样加药、肝匀浆加药及肝匀浆合并空白血加药温孵16 小时,当血液由鲜红转为暗红,且发绿时,代谢转化率最快。(9)莱格注射液对抗甘油所致急性肾功能衰竭的实验表明,莱格注射液可明显降低血清肌酐浓度,尤以给药 3 天后作用最明显。结论(1) 首次将莱格内酯制成了安全、有效、可控、稳定的静脉脂肪乳-莱格注射液,解决了该成分难于稳定保存这一技术难题。(2) 首次基本全面的研究了莱格内酯注射液在SD大鼠和Beagle犬的体内药物代谢动力学特点,该药的体内药时行为符合三室模型,粪、尿和胆汁的原形排泄总量不到10%率。体内的代谢产物正在研究中。(3) 莱格内酯具有很强的蛋白结合力,人血浆蛋白结合率在 97%以上(血浆来源为 301 医院输血科采集的献血战士的混合血浆)。主要创新点⑴ 以脂肪乳为载体,首次将莱格内酯制备成了脂肪乳注射液,解决了该成分难于稳定保存这一技术难题;针对莱格内酯这一方面的研究国内乃至国际上尚未见报道。⑵ 首次基本全面的研究了莱格内酯注射液的药物代谢动力学特点,为该药的临床使用提供了可供借鉴的药动学参数。 主题词急性肾功能衰竭;莱格内酯;生物转化;药物代谢动力学;脂肪乳
OBJECTIVE: Lig Lactone shows very strong activity in cure Acute Renal Failure without correlated medicine report in the world.It’s raw material was extracting from plant.The intravenous emulsion was prepared with pressure homogenizing technology(HE).The preparation is belong to the first species new medicine of traditional Chinese medicine.We extract ed Lig Lactone raw material. The Reverse-phase HPLC method was established to control the emulsion’s quality.The stability was studied to develop a useful method for it’s preservation.The pharmacodynamics was investigated to provide the bases for clinical application. Plasma protein binding ratio was determine to know the drug is binding-sensitive. METHODS The intravenous emulsion was preparation with high pressure homogenizing technology(HE). The Reverse-phase HPLC method with ultraviolet detection was used to determine the content of Lig Lactone and the variety of the content when lighted and heated and accelerated. After the injection of the intravenous emulsion of LIG within rats caudal vein and dog forelimb vein, we assay the concentration of Lig Lactone in blood vs time and use software 3p87 calculate the data of concentration ,so that we can make sure the rudiment procedure and the pharmacokinetics of Lig Lactone in vivo. After the injection of the intravenous emulsion of LIG within mouse caudal vein,we assay the concentration of Lig Lactone in different organs and blood at different time. After the injection of the intravenous emulsion of LIG, we assay the concentration of Lig Lactone in stool and urine and bile to calculate the total excretion amount of it. Through the test that liver homogenization and blood metabolize Lig Lacton in vitro. ,we may deduce the possible metabolism route of the Lig Lactone. Use ultrafiltration determine the plasma protein binding ratio RESULTS (1)The established reversed HPLC method is reliable. The stability and precision and accuracy specificity and reproducibility and toleration of the method are qualified.The regression equation is y=18.931x-3.2601,r2=0.9999(relation betweent he peak area of Lig Lactone and the sample size by HPLC-UV method) (2)The results of accelerated tsetse showed that the colur and the content of Lig Lactone and the mean diameter and PH were kept still under 25℃ after 9 months ,but the content droped slightly after 12 month . The results of accelerated tsetse showed that the content of Lig Lactone droped gradually under 40℃ in 1 to 6 months. .The influential factors proved that the light can affect remarkably the content of Lig Lactone of the emulsion and the content of Lig Lactone dropped 60% under 4700Lx after 10 days.The long-term tests proved that the contentand the colur and the mean diameter and PH are stabile under 6℃ in commercially available back. The intravenous emulsion shoud be stored under 6℃±2℃. (3)The results that the c-t date that the intravenous emulsion was injected within rats caudal vein (the dose is 50mg/kg、20mg/kgand5mg/kg) analyzed by 3p87 computing program showed that the pharmacokinetic behavior of the intravenous emulsion of lig fitted a three-compartment open pharmacokinetic model. The distribution phase half-life,t1/2pi(50mg/kg、20mg/kgand5mg/kg) is respectively 8min、2 min and 4min .t1/2α(50mg/kg、20mg/kg and 5mg/kg )is respectively 27min、23 min and20 min.The elimination phase half-life, t1/2β(50mg/kg、20mg/kg and 5mg/kg)is respectively 330min、421min and 618 min .AUC of these doses (50mg/kg、20mg/kg and 5mg/kg) which were caculated by statistic moment algorithm are respectively 2811.5and1695.2and 755.7(μg/ml )(min). (4) The results that the c-t date that the intravenous emulsion was injected within rats caudal vein (the dose is 10mg/kg) in multiple-dosage showed that the pharmacokinetic behavior of the intravenous emulsion of lig fitted three –conpartment model. The distribution phase half-life of the first dosage and the last dosage,t1/2pi is respectively 3.6min、8.6 min,t1/2αis respectively 25min、13min and20min. (5) The results that the c
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