五味风湿胶囊新药研究

【作者】 王秀兰；

【导师】 杨松松；

【作者基本信息】 辽宁中医药大学 ， 生药学， 2006， 博士

【摘要】 蒙药忠伦-5汤由苦参、栀子、诃子、川楝子和地格达组成。具有清热，凉血之功效。可以收敛、杀击陈旧性热，隐伏热，治疗热性协日乌苏病，痛风，痹病等。目前，蒙医临床上主要用于协日乌苏引起的关节病及皮肤病。 本课题将传统蒙药验方忠伦-5汤研制成五味风湿胶囊。确定了五味风湿胶囊最佳提取工艺；建立了五味风湿胶囊质量控制标准；同时对其进行了抗炎镇痛、抗类风湿药效研究及急性毒性研究。 运用正交实验法研究了最佳提取工艺，苦参提取工艺以苦参碱和氧化苦参碱为对照品，其它四味药以栀子苷为指标，用高效液相色谱（HPLC）法进行含量测定。实验结果表明，苦参提取最佳工艺为A₁B₃C₂D-2，即用8倍量的60％乙醇回流2次，每次回流1.5h。其它四味药最佳提取工艺为A₂B₁C₂，即加8倍量水煎煮2次，每次煎煮1h。 利用薄层色谱法对苦参、栀子、诃子及紫花地丁进行定性鉴别，运用HPLC法对苦参碱、氧化苦参碱及栀子苷进行含量测定。实验结果表明，薄层色谱中各味药的特征斑点清晰，重现性好。含量测定方法精密度、稳定性及重复性良好。苦参碱+氧化苦参碱及栀子苷回收率分别为101.2％、98.32％，RSD分别为2.15％、2.1％。 对五味风湿胶囊进行装量差异，水分测定，崩解时限等一般检查。一般检查结果全部符合中华人民共和国药典（2005版）有关要求。稳定性实验结果表明，室温放置12个月各种指标稳定。 通过二甲苯致小鼠耳廓肿胀试验、角叉菜胶致大鼠足肿胀试验、棉球致大鼠肉芽肿试验、Ⅱ型胶原性小鼠关节炎试验、佐剂性关节炎（AA）试验及AA大鼠脾细胞培养、AA大鼠腹腔巨噬细胞培养液对C57BL／6J小鼠胸腺细胞增殖能力的影响等试验，考察了该药主要药效。结果表明，五味风湿胶囊对二甲苯致小鼠耳廓肿胀、角叉菜胶致大鼠足肿胀、棉球致大鼠肉芽肿均有明显的抑制作用。对AA大鼠原发性、继发性关节炎及Ⅱ型胶原性小鼠关节炎均有显著的抑制作用；五味风湿胶囊可降低AA大鼠关节组织中白介素-1β、肿瘤坏死因子-α、一氧化氮（NO）及前列腺素（PGE）水平；抑制AA大鼠脾细胞增殖能力；抑制C57BL／6J小鼠胸腺的增殖能力；显著提高血清超氧化物歧化酶（SOD）活性。小鼠急性毒性试验结果表明，五味风湿胶囊LD₅₀大于98.37g（生药）／kg。 实验设计和操作过程中充分运用现代科学手段的同时注意保留蒙药传统用法特色。本课题对五味风湿胶囊的进一步研究提供了实验依据。同时对传统蒙药的进一步开发研究和推广应用具有重要的参考意义。更多还原

【Abstract】 Mongolian drug Zhonglun-5 decoction is composed of Sophora flavescens Ait, Gardenia jasminoides Ellis, Terminalia chebula Retz, Viola yedoensis Makino. It is provided with the effect of heat clearing, removing heat from the blood. It can astringe and resist old heat, hidden heat, and cure feverish xiriwusu disease, severe migratory arthralgia, stagnation-symdrome of Qi and blood and the like. At present, it is mainly used in arthropathy and dermopathy caused by xiriwusu in clinic of Mongolian medicine.The traditional Mongolian drug zhonglun-5 decoction was developed into wuweifengshi capsules in this study. The best extractive technology is determined. The qualitative control criterion of wuweifengshi capsules is set up, and at the same time a study of its anti-inflammatory, abirritation, anti-rheumatoid arthritis action was made.A study of the best extractive technology was made with orthogonal design. The extractive technology of Sophora flavescens Ait used matrine and oxymatrine as the contrastive object while the other four drugs used gardenoside to test the content with HPLC. The experimental results showed the best extractive technology of Sophora flavescens Ait. was A1B3C2D2, namely eight times of 60% ethanol in quantity was decocted twice, ninety minutes each time. The best extractive technology of the other four drugs was A₂B₁C₂, namely eight times of water in quantity was added, decocted twice, sixty minutes each time.A qualitative identification of Sophora flavescens Ait, Gardenia jasminoides Ellis, Terminalia chebula Retz, Viola yedoensis Makino was made with thin layer chromatogram. The content of matrine and oxymatrine was tested with HPLC. The experimental results showed that the characteristic spots of each drug in the chromatogram of thin layer are clear and distinct. The reappearance is good. The content determination method is practical, recovery of matrine plus oxymatrine, gardenoside were 101.2% and 98.32%, RSD更多还原