【作者】 陈武；

【导师】 刘志辉；

【作者基本信息】 南京中医药大学 ， 药剂学， 2011， 硕士

【摘要】 卵巢功能早衰(Premature ovarian failure, POF)是指妇女在40岁以前因某种原因引起的闭经、不育、雌激素缺乏、以及促性腺激素水平升高为特征的一种疾病,是由于卵巢合成性激素功能低下,或者不能合成,降低了对下丘脑一垂体轴的负反馈作用,使得促生殖激素增高,雌激素水平降低的一种状态。其主要特点是卵巢功能、生殖功能和内分泌功能丧失。目前,对于卵巢功能早衰尚无肯定有效的治疗方法。西医治疗POF的方法均有不同程度的副作用。患者中除少数患者通过雌激素替代治疗或免疫抑制法,可恢复和改善卵巢功能,多数患者尚无理想的治疗方法；中医药治疗POF具有独特的优势。乌鳖颗粒处方为江苏省中医院名老中医夏桂成教授的经验方,由制何首乌、白术(麸炒)、续断、枸杞子、茯苓、鳖甲(炙)和黄狗肾共七味中药组成,用于治疗卵巢功能早衰(POF),临床应用多年,疗效显著。本课题按照国家中药新药研究指导原则对该处方进行了药学文献研究、制剂工艺研究、质量标准研究以及稳定性考察。药学文献研究部分对处方中七味药的来源、性状、化学成分、定性与定量分析、药理作用等方面进行了相关文献研究,并进行了综述总结。制剂工艺研究部分1.根据中医药理论、各药味主要有效成分的性质设计制剂工艺路线并对工艺路线进行了药效学筛选；2.以相关药效学评价为指标,优选黄狗肾的提取工艺条件为：加10倍量95%乙醇提取2次,每次1h；3.对黄狗肾进行了相关探索研究,采用超临界CO2流体萃取黄狗肾药效成分并对萃取物进行GC-MS分析,结果优选的萃取工艺条件为：萃取压力25MPa,萃取温度65℃；分离釜Ⅰ压力5MPa,温度40℃；分离釜Ⅱ压力5MPa,温度35℃；夹带剂为1倍量95%乙醇；萃取时间2h,萃取物得率为27.4%。共鉴定出44种脂肪酸成分,主要为8-十八碳烯酸、14-甲基十五烷酸、16-甲基十七烷酸及硬脂酸等；同时,对黄狗肾不同提取物进行药效学比较研究及性激素含量测定,结果醇提物和SFE-CO2提取物药效强于水提物。三种提取物TESTO、E2含量则相差不大,均以SFE-CO2提取物最高,其次为醇提物。4.采用正交试验以多糖含量和氮含量为指标优选了鳖甲(炙)、枸杞子、茯苓煎煮工艺条件：加8倍量水煎煮3次,每次60min；5.采用单因素考察和正交试验相结合以二苯乙烯苷和川续断皂苷Ⅵ为指标优选了制何首乌、续断、白术(麸炒)煎煮工艺条件：制何首乌粉碎成20目粗粉,与续断、白术(麸炒)饮片加8倍量水提取3次,每次60min；并通过正交试验优选了醇沉除杂工艺条件：水提液浓缩至相对密度1.15(60℃测),加乙醇至含醇量60%,室温静置12h；6.考察了浓缩和干燥的方式、温度等对效果的影响,优选浓缩和干燥工艺条件为：温度70-75℃,真空度0.05-0.08 MPa条件下进行浓缩；减压干燥时条件控制在温度不超过70℃,真空度0.08-0.10MPa；7.在成型工艺中,考察了辅料种类和用量,结果以50%(占制剂的百分比)的糊精和0.2%的阿司帕坦作为辅料,采用一步制粒并优选了制粒工艺参数：提取液相对密度1.10-1.15(60℃测),进料速度20-25rpm。雾化压力0.15-0.25MPa,进风温度80-90℃,开始阶段进风量80-90m3/h,最后进风量控制在150-180m3/h进行干燥；8.按照优选的工艺条件进行了三批中试研究,结果表明优选的工艺条件稳定、可行,具有可操作性。质量标准研究部分1.对三批中试产品进行了制剂通则检查,结果符合规定；2.采用薄层色谱法,以对照品和(或)对照药材为对照,建立了制剂中制何首乌、白术(麸炒)、续断、枸杞子、茯苓的定性鉴别方法；3.采用HPLC法对制剂中的二苯乙烯苷和川续断皂苷Ⅵ两种活性成分进行了同时测定。并制定了相应的含量限度；4.采用HPLC法建立了制剂的指纹图谱,方法稳定、可靠、重现性好,为制剂的质量控制和评价提供了依据。稳定性考察部分对三批中试产品进行了3个月的加速试验考察。结果表明制剂在考察期间稳定性良好。更多还原

【Abstract】 Premature Ovarian Failure(POF) is a disease caused by some factor,it characteristiced with amenorrhea、infertility、estrogen anhormonia and high GTH in those women befor the age of 40.It is a state that ovarian gonadal hormone hypofrontality or disability decreases the negative feedback to hypothalamic-pituitary axis,then leads the increase of GTH and the decrease of estrogen.The main feature of POF is disabilities of ovary、reproduction and endocrine.There are no effective therapies for POF recently,Western medicine have some serious side-effects,few of them can improve or even recover ovarian functions by hormone replacement therapy or immunodepression,There are still lack of desirable therapies for most patients.Traditional Chinese Medicine have its unique advantages in therapies for POF.The prescription of Wubie granules is a proved effective recipe that has been used for many years in the treatment for POF by the famous doctor of Traditional Chinese Medicine——Xia Gui-cheng.The recipe is made up of Radix Polygoni Multiflori Preparata、Rhizoma Atractylodis Macrocephalae、Radix Dipsaci、Fructus Lycii、Poria、Carapax Trionycis and Testis et Penis Canitis.This project is to study related literature、preparation technology、quality standard and stability of Wubie granules according to the demands of new preparations for Traditional Chinese Medicine.Literature Section:Each medicine in the recipe has been summarized,including development in source、characteristics、chemical constituent、qualitation and quantitative analysis and pharmacologic actions.Preparation Section:1.According to the theory of Traditional Chinese Medicine and the physicochemical property. of the active ingredient, preparation process has been designed and evaluated by pharmacodynamics indexes; 2.The optimum extraction process of Testis et Penis Canitis evaluated by pharmacodynamics indexes were as follows:extracting 2 times, using 10-fold of 95% alcohol amount for 1 h each time; 3.For further research, the optimum parameters of the supercritical CO2 fluid extraction of Testis et Penis Canitis and chemical constituents of extraction have been studied by GC-MS:the extraction pressure was 25MPa and temperature was at 65℃,1mL/g of 95% alcohol as entrainer, extracted for 2 h and the extractive Rate was 27.4%;44 types of fatty acids were identified from the extraction. Meanwhile,3 types of Testis et Penis Canitis extraction have been evaluated by pharmacodynamics and sex hormone was determined. The results are:the alcohol extraction and the SFE- CO2 extraction are superior to the water extraction. The concentration of TESTO and E2 in the 3 types of extraction are proximate, the SFE- CO2 extraction is highest,and the alcohol extraction followed; 4. Orthogonal design was used to optimize extracting process of Fructus Lycii、Poria and Carapax Trionycis,using the content of polysaccharide and nitrogen as indexes,and the results are: extracting 3 times, using 8-fold of water amount for 60 minutes each time; 5. Single factor and orthogonal design were used to optimize extracting process of Radix Polygoni Multiflori Preparata、Rhizoma Atractylodis Macrocephalae and Radix Dipsaci,The content of tetrahydroxystilbene-glucoside and asperosaponin VI were indexes. and the results are:Radix Polygoni Multiflori Preparata is shattered to coarse powder,then extracting 3 times with the others, using 8-fold of water amount for 60 minutes each time;Then orthogonal design was used to optimize the alcohol precipitation process of the water extraction,The results are:the relative density of the extraction is 1.15(determined under 60℃), adding alcohol to the extraction and making it to 60% alcohol solution, then letting it stand for 12 hours at room temperature; 6. Concentration and drying process were studied, The results are:the extraction is concentrated with 70-75℃in temperature,and the vacuum is 0.05-0.08 MPa;the temperature is 70℃,and the vacuum is 0.08-0.10MPa while drying; 7. Preparation process for the granules has been optimized; 8.Pilot scale experiments have been carried out using the studied preparation process.Quality Standard Section:1. According to the Chinese Pharmacopoeia (edition 2010) general principles for granules,the pilot scale products have been checked; 2. The identification methods of the medicines in the preparation have been established using TLC; 3. The simultaneous Determination method of tetrahydroxystilbene-glucoside and asperosaponin VI has been established using HPLC; 4. The method of finger print for the preparation has been established using HPLC.Stability Section:The accelerated test of the preparation has been carried out for 3 months,and the results show that it is stable in the test period.更多还原