去氢厄弗酚缓释片的制备工艺及体内外释药研究

【作者】 高伟；

【导师】 黄建梅；

【作者基本信息】 北京中医药大学 ， 中药分析学， 2018， 硕士

【摘要】 研究背景和目的:去氢厄弗酚是从中药灯心草中分离得到的一种菲类化合物,课题组前期通过小鼠高架十字迷宫实验和洞板实验,证实去氢厄弗酚具有显著的抗焦虑作用,是灯心草清心除烦的物质基础之一。对去氢厄弗酚在小鼠体内的药代动力学和体内过程进行研究,达峰时间5-10min左右,消除半衰期200-260min左右,其抗焦虑作用的靶器官可能是脑组织,在抗焦虑剂量下无肌松副作用。基于前期研究,本课题拟将去氢厄弗酚制备成缓释片,进行体外体内释药研究,为一类新药去氢厄弗酚的临床前提供基础。方法:1以灯心草干燥茎髓为原料,硅胶柱色谱与薄层色谱(TLC)结合,制备纯化去氢厄弗酚,通过核磁共振波谱(NMR)定性,高效液相色谱(HPLC)定量,得到纯度>90%的原料药。2以HPMC作为骨架材料,制成去氢厄弗酚亲水凝胶骨架型缓释片。以去氢厄弗酚为体外释放指标,测定药物的累积释放度,考察HPMC、填充剂及润滑剂等处方因素对去氢厄弗酚释放的影响,设计正交试验筛选最优处方并进行验证。3建立UV测定去氢厄弗酚含量的检测方法,以去氢厄弗酚为体外释放指标,考察释放介质、温度、体积,试验溶出装置、转速及仪器等因素对药物释放的影响;对去氢厄弗酚缓释片进行释药曲线拟合及机理阐释。4采用HPLC测定去氢厄弗酚的血药浓度,以自制去氢厄弗酚普通片作参比制剂,自制去氢厄弗酚缓释片为受试制剂进行兔体内药动学研究和体外相关性评价。结果:1得到纯度>90%的去氢厄弗酚单体20 g。2得到的去氢厄弗酚缓释片最优处方为:HPMC K4M用量为35%,MCC用量为15%,乳糖用量为44%,硬脂酸镁为1%,全粉末直接压片。所制备的缓释片2 h、6 h、12 h的累积释放率为30%、60%、90%,重现性与均一性良好。3建立的体外释放度测定条件为:0.5%吐温-80溶液900 mL为释放介质,温度37±0.5℃,桨法,转速100rpm。释放介质的pH、溶出仪器等对去氢厄弗酚的释放无影响。去氢厄弗酚的释药曲线可用一级动力学方程拟合,释药机理为药物扩散与凝胶溶蚀协同作用。4兔口服自制去氢厄弗酚缓释片和普通片后,血药浓度经药动学处理软件DAS 2.0进行药物动力学参数拟合,符合二室模型,两者的Cmax分别为3.99、12.16 ng·mL-1;tmax分别为 3、2 h;AUC0-∞分别为 17.692、22.029 ng·h·mL-1;选用 Loo-Riegelman 方程,证明去氢厄弗酚缓释片体内外释药相关性良好。结论:通过硅胶柱色谱与薄层色谱(TLC)结合,可从灯心草干燥茎髓中提取分离得到去氢厄弗酚单体,得率约为0.1%。制备的去氢厄弗酚缓释片前期无突释,中期释药平稳,后期释药完全,达到了 12 h的缓释效果,其重现性与均一性良好。去氢厄弗酚缓释片体外释药曲线为一级动力学方程,释药机理为扩散与溶蚀协同作用。兔口服缓释片后,与普通片相比,Cmax减少,tmax延长,MRT延长,显示明显的缓释效果;去氢厄弗酚缓释片体内外释药具有良好的相关性。更多还原

【Abstract】 Background and objective:Dehydroeffusol is a phenanthrene compound,which was isolated from Junsus effusus L.of traditional Chinese medicine.It has an anxiolytic effect was confirmed through the elevated plus-maze test and hole-board test in mice,which was one of the active constituents of restlessness and insomnia in Junsus effusus L,in our previous experiments.The pharmacokinetics and in vivo processes of dehydro effusol in mice were studied,and the elimination half-life of about 200～260 min,the target organ for anxiolytic effect may be brain tissue,and no muscle relaxation side effect under anxiolytic dose.Based on the previous studies,this project intends to prepare dehydroeffusol sustained-release tablets for in-vitro studies in vitro,providing a basis for the clinical application of a new class of drugs for the dehydroeffusol.Method:1 The dried medulla common rush was used as the raw material,and silica gel column chromatography was combined with thin layer chromatography(TLC)to prepare and purify the dehydroeffusol.Qualitative analysis was performed by nuclear magnetic resonance spectroscopy(NMR)and the purity was determined by high performance liquid chromatography(HPLC).2 HPMC as a backbone material was used to prepare a dehydroeffusol hydrophilic gel matrix sustained-release tablet.The release of dehydroeffusol was used as an in vitro release index,using the ultraviolet and visible spectrophotometry(UV)to determine the cumulative release of the drug,the effects of prescribing factors such as HPMC,fillers and lubricants on the release of dehydroeffusol were investigated.The orthogonal test was designed to optimize prescription and verify.3 The dehydroeffusol was used as an in vitro release indicator,the influence of the release medium,temperature,volume,and the the dissolution apparatus,rotation speed,and instrumentation on the release of the drug were investigated.Dehydroeffusol sustained-release tablets for drug release curve were fitted and release mechanism was elucided.4 A HPLC method was used for the determination of the concentration of dehydroeffusol in blood.The self-made dehydroeffusol tablet was used as a reference preparation,the dehydroeffusol sustained release tablet was used as a test preparation to study the pharmacokinetics in rabbits and in vitro correlation assessment.Result:1 Obtained>90%purity of dehydroeffusol monomer 20 g.2 The optimal formulation of the dehydroeffusol sustained-release tablets was:HPMC K4M 35%,MCC 15%,lactose 44%,and magnesium stearate 1%and prepared by whole powder was directly tableted.The cumulative release rate of the sustained-release tablets at 2 h,6 h,and 12 h was 30%,60%,and 90%,and the reproducibility and homogeneity were good.3 The established in vitro release assay conditions were:0.5%Tween-80 solution 900 mL as release medium,temperature at 37±0.5℃,paddle method,speed at 100 rpm.The pH of the release medium,the dissolution apparatus,etc.have no effect on the release of dehydroeffusol.The release profile of dehydroeffusol can be fitted by the first-order kinetic equation.The drug release mechanism is synergistic between drug diffusion and gel erosion.4 The rabbits oral administrationed self-made dehydroeffusol sustained-release tablets and ordinary tablets,the blood drug concentration by pharmacokinetic processing software DAS2.0 for pharmacokinetic parameter fitting,the result showed that the absorbtion of dehydroeffusol in rabbits conforms two-bedroom model.The Cmax of the two was 3.99 and 12.16 ng·mL-1,respectively;the tmax was 3 and 2 h,and the AUC0-∞ was 17.692 and 15.164 ng·h·mL-1,respectively.The Loo-Riegelman method was good in in vitro and in vivo.Conclusion:By combining silica gel column chromatography with thin layer chromatography(TLC),dehydroeffusol monomers can be extracted and separated from dried medulla common rush with a yield of approximately 0.1%.The prepared dehydroeffusol sustained-release tablets have no burst release in the early stage,stable release in the middle stage,complete release of the drug in the later stage,reaching a sustained release effect of 12 hours,and good reproducibility and uniformity.The in vitro release profile of dehydroeffusol sustained-release tablets is a first-order kinetic equation.The mechanism of drug release is the synergy between diffusion and dissolution.Compared with ordinary tablets,the amount of Cmax in dehydroeffusol sustained release tablets decreased and tmax extended,which showed obvious release effect,used the Loo-Riegelman method to do correlation between in vivo evaluation and found that the Sustained release tablets in vivo correlation is more significant.更多还原