【作者】 罗晓健；

【导师】 张汝华；

【作者基本信息】 沈阳药科大学 ， 药剂学， 2001， 博士

【摘要】 中药制剂是祖国医药学的重要组成部分，是我国劳动人民长期医疗实践中的经验总结，为中华民族的繁衍昌盛发挥了重要作用。但与西药制剂相比，存在工艺落后，剂型陈旧，缺乏科学评价制剂质量的客观指标和制剂安全有效的科学数据，中药制剂基础研究薄弱，新剂型研究开发能力差等问题，严重阻碍中医药事业发展和中医药走向世界，因此以中医药理论为指导，采用现代科学技术，对中药制剂进行现代化研究具有十分重要的意义。 复方丹参片为《中国药典》2000年版一部收载品种，由丹参、三七、冰片组成，具有活血化瘀，理气止痛之功能，用于治疗冠心病、心绞痛。其疗效确切，临床应用广泛，方中药味较少，基础研究较深入，本文选择复方丹参片作为中药复方缓释制剂的模型药物，主要进行了以下研究工作，取得了较满意的结果。 1．采用TLC方法对复方丹参多层缓释片中丹参素、原儿茶醛、人参皂苷Rg₁、Rb₁、冰片进行了鉴别。首次采用LC／MSⁿ技术对三七、丹参提取物和复方丹参缓释片溶出液进行了组分分析；采用HPLC法对丹参素、原儿茶，人参皂苷Rg₁进行了含量测定；采用GC法测定冰片的含量；首次采用HPLC法用荧光检测器测定犬体内丹参素血药浓度。以上方法灵敏准确，简单实用，为本课题的研究提供了可靠的检测手段。 2．考察了影响指标性成分提取率的工艺因素，并采用均匀设计优化了提取工艺。用水提醇沉法提取丹参酚酸类成分，丹参素含量大于5.0％，原儿茶醛含量大于0.5％；对大孔吸附树脂吸附三七总皂苷的吸附性能进行了系统研究，采用稀醇提取、大孔吸附树脂纯化三七总皂苷，总皂苷含量大于85％（以人参皂苷Rg₁计）。用LC／MSⁿ技术鉴别出三七提取物含有人参皂苷Rg₁、Rg₂、Rb₁、Rd、Re、三七皂苷R₁、R₂、R₃、R₄，丹参提取物含有丹参素、原儿茶醛、原儿茶酸、丹酚酸A、B、C、E、F，均与三七、丹参原药材所含的主要成分一致。药效学实验结果显示，复方丹参提取物（由丹参提取物、三七提取物、冰片按适当比例组成）可显著增加豚鼠离体心脏冠脉流量，对垂体后叶素所致大鼠心肌缺血有很好的保护作用，其效果明显优于复方丹参片…＜005人与复方丹参滴丸无显著差异中＞0刀5人 说明复方丹参提取物基本保留了药材及处方中主要有效成分，提取工艺合理可行。 3 对丹参提取物、三七提取物、冰片制剂处方设计前的主要理化性质进行了测试。结果表明，丹参提取物、三七提取物易溶于水，冰片难溶于水，三七提取物可增加冰片在水中的溶解度。丹参提取物、三七提取物、冰片流动性差，丹参提取物、三七提取物中指标性成分对光、热、湿稳定，丹参提取物吸湿性强，冰片易升华挥发。首次对复方丹参提取物中丹参素、原儿茶醉、及冰片的大鼠在体小肠吸收动力学进行了研究，结果显示指标性成分在大鼠小肠主要通过被动转运机制吸收，符合为一级吸收过程，胆管结扎与否和回流液pH值在3刀刃4的范围内，对指标性成分吸收均无明显影响。这些工作为剂型的选择和制备工艺的确定提供了依据。 4．复方丹参多层缓释片由速释层、冰片三七缓释层、丹参缓释层组成，以保证制剂中药物同步释放，快速起效，持续作用。速释层以PVPP为崩解剂，MCC、乳糖作填充剂，采用混料设计试验方法优化处方，制成的速释层在Zmin内崩解；冰片三七缓释层以 HPMC为主要骨架材料，用 L－HPC、PVPK3 0调节药物释放速度，经处方优化后，指标性成分冰片、人参皂苦Rgl的释放速度符合要求。以丹参素为指标性成分，考察了EC的粘度、用量、润滑剂、阻滞剂和压力等因素对释药速度的影响，并用正交设计优化了处方和工艺，制备的丹参缓释层具有较好的缓释作用。首次详细研究了多层片压制时加料顺序和压力对丹参素和冰片释放的影响，结果表明，释药层在多层片中位置相同时，压力对指标成分释放速度无明显影响；压力相同时，释药层的位置对指标成分的释放影响较大，特别是在中间层位置可明显降低多层片药物的释放速度。复方丹参多层缓释片最佳压制工艺为56．7MPa压力，三层加料顺序为速释层、冰片三七缓释层、丹参缓释层。 5．由于中药复方成分复杂，如何科学评价中药缓释制剂体外释药过程是一个急待解决的问题。本文首次采用LC／MS”技术对中药缓释制剂释放浪进行组分分析和用HPLC测定溶出液的指纹图谱，以考察缓释层中主要成分是否能够溶出，寻找指纹峰与参照峰的相关性，试图阐明中药复方缓释制剂多种成分的释放 2行为和机制。研究结果显示，丹参缓释层、冰片三七缓释层释放液（6h）含有人参皂昔 Rgl、RgZ、Rbl、Rd、RC、三七皂苦儿、RZ、R3、凡、丹参素、原地茶醛、原儿茶酸、丹酚酸A、B、C、E、下，与三七、丹参提取物所含主要成分一致，提示己知主要成分均可从多层缓释片中按设计要求释放出来。在HPLC指纹图谱中，选择原儿茶醛为参照峰，算出其它指纹峰与原儿茶醛峰面积的线性相关方程行均川95人 根据原儿茶醛的标准曲线方程，求出其它指纹峰成分相对释更多还原

【Abstract】 Traditional Chinese Medicine Preparations (TCMP) is an important part of Traditional Chinese Medicines (TCM) which is the sum of medicine experience of the labouring people in long time and has played a great role in the prosperity of Chinese nation because of their precise composition, good curative effects and little side-effects. However, TCMP is faced with many problems such as outdated dosage forms, backward techniques, which have hindered the development of TCM undertakings and movement toward the world. It is urgent to study TCMP by making full use of advanced science and techniques.Compound Danshen tablets (CDT), listed in the first section of the 2000’s edition of "Pharmacopoeia of the People’s Republic of China", consists of the extract of radix Salvia miltiorrhiza (Bunge), root ofPanax notoginseng (Burk), borneolum. It is primarily applied to the treatment of coronary artery disease, angina pectoris. In this thesis, CDT was selected as the model drug of Traditional Chinese Medicine compound multi-layer sustained release tablets (TCMCSRT) because of only three herbs in the recipe, more extensive basic researches and good curative effects. The main researches made and results were as follows.1. TLC methods were used to identify the Danshensu (DS), protocatehuic aldehyde (PA), ginsenoside Rgi, Rbi, and borneol. The methods of UV for total panax notogiseng saponins (PNS). The HPLC for SD, PA and ginsenoside Rgi and the GC for borneol were developed to detect their contents in the preparations. The HPLC method with fluorescence detection was the first analytical method for the determination of DS in dog plasma. Above-mentioned analytical methods were simple, accurate and suitable for in future study of compound Danshen multi-Layer sustained release tablets (CDMSRT).2. The factors such as heating temperature, decoction time, alcohol concentration, which was influenced on the extraction and purification of target constituents were investigated. The extraction processes of radix Salvia miltiorrhiza and root Panaxnotoginseng were optimized using the uniform design respectively. The extraction of Salvia miltiorrhiza (ESM) was obtained by boiling the crude herb with water and precipitating with alcohol, whose DS content was 5.7%. Extracts of Panax notoginseng (EPN) was drawn with 30% alcohol and purified with macroporous resin D101, the total saponins content was 85.0%. Ginsenoside Rg1, Rg2, Rb1, Rd1, Re, notoginsenoside R1, R2, R3, R4, DS, PA, protocatechuic acid, salvianolic acid A, B, C, D, E, F were differentiated from the extracts through LC/MSn. It implied that there was no marked difference in the kinds of main constituents between crude herbs and the extracts. The study on the pharmacodynamics indicated that compound Danshen extracts (CDE) could evidently increased isolated guinea pig ardiac coronary artery blood flow (P < 0.05) and nullified the pituitrin-induced electrocardiographic ischemic ST-T (P < 0.05) as compared with control group. But there was no significant difference (P > 0.05) between CDE and Compound Dropping Pills. It seemed that the majority of effective constituents of CDT were still retained in the CDE, and the preparation processes for CDE were proper and practicable.3. The study on physicochemical properties of compound Danshen powders (CDP) showed that ESM and EPN were hydrophilic. Borneol was hydrophobic, whose solubility could be increased when it was compatible with EPN. Treatment with the light, temperature and moisture, the content of DS, PA and Rg1 in ESM and EPN were kept unchanged. The flowbility of CDP was poor. EDS was hygroscopic. Borneol possesed sublimation. Intestinal absorption experiments of rats in situ were used to study the absorption of DS, PA and borneol in CDP for the first time. It was found that the absorption of DS, PA and borneol complied with the first order kinetics by passive transport mechanism and was obviously influenced by pH value of regurgitation fluid, with and without ligation of rat’s bile duct. There were no更多还原