栀子抗抑郁活性部位微球的制备工艺研究
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摘要
栀子是茜草科植物山栀子的干燥成熟果实,性寒、味苦,无毒,具有泻火除烦,清热利湿,凉血散瘀的功效。该药是2010年版《中国药典》收载品种,主要用于热病心烦,急性黄疸型肝炎,膀胱炎,目赤肿痛的治疗。目前对栀子的药理研究较多,主要是集中在抗哮喘作用、抗炎镇痛作用、利胆保肝作用、抗病毒作用及对神经系统的作用等。我们前期试验证明栀子乙醇提取物具有抗抑郁活性;栀子乙醇提取物中有效成分为栀子苷,但由于栀子苷存在半衰期短的缺点,所以本课题采用乙醇回流提取栀子苷,采用大孔吸附树脂进行纯化,并通过药效学实验验证栀子提取物的抗抑郁作用,利用喷雾干燥法将其制备成微球,从而延长其半衰期,提高生物利用度,更好地发挥其疗效。
目的:研究栀子抗抑郁活性部位的最佳提取、纯化工艺,并将其制备成微球。
方法:通过小鼠抑郁模型,以累计游泳不动时间为考察指标,筛选栀子的抗抑郁活性部位。采用L_9(3~4)正交试验法,以出膏率和栀子苷的含量为考察指标,HPLC法测定栀子苷的含量,优选栀子最佳提取工艺;采用大孔吸附树脂法对栀子的粗提取物进行纯化,通过对大孔吸附树脂的型号、吸附速度、吸附时间、洗脱剂浓度、洗脱速度及洗脱剂用量等因素的考察,确定最佳纯化工艺;采用喷雾干燥法制备微球,通过单因素试验法,确定最佳载体材料;通过正交试验法,考察药物与载体材料比、进风温度、出风温度、进料速度,以收率、包封产率为评价指标,确定微球的最佳制备工艺;同时对微球的形态、粒径及其分布、突释效应进行检查,通过测定其体外释放度,进行栀子抗抑郁活性部位微球释药机理的初步研究。
结果:栀子60%乙醇提取物具有明显的抗抑郁作用;栀子最佳提取工艺为加6倍量60%的乙醇,提取3次,每次1h;栀子苷的纯化工艺为AB-8型大孔吸附树脂,以2ml/min的速度上样2倍柱体积0.25g生药/ml的栀子提取液,吸附1h,先用5倍柱体积的水洗脱杂质,再用6倍柱体积的40%乙醇以3ml/min的速度进行洗脱;采用喷雾干燥法制备微球,最佳工艺:药物与载体材料比为1:2,进风温度为130℃,出风温度为90℃,进料速度为300ml/h;结果显示栀子抗抑郁活性部位微球中栀子苷的释药行为符合Higuchi模型。
结论:工艺简单合理、适合大工业生产。
Fructus gardenia is the dry ripe fruit of Mountain gardenia Rubiaceae, cold-natured, bitter in taste, innocuity, with the function of reducing fire except vexed, clearing away heat evil and promoting diuresis, cooling blood eliminating stasis to activate blood circulation. The drug is one piece of the 2010 edition of China pharmacopoeia, it is mainly used for treat fever upset, acute icteric hepatitis, cystitis, conjunctival congestion score pain, upper gastrointestinal hemorrhage and partly to bleed, bruised. At present the pharmacological research of fructus gardenia is increasing, main is concentrated in resistance to asthma action, anti-inflammatory analgesic action, cholaneresis hepatoprotective effect, antiviral effect and the function of nervous system etc. The early experiment proofed the ethanol extract of fructus gardenia has antidepressant activities. However ,the geniposide which is the active of fructus gardenia the feature of the short of half-life.So this subject adopts ethanol refluxing extraction geniposide, uses the macroporous adsorption resin in purification, And through the pharmacodynamics experiment verify the extract of fructus gardenia antidepressant effect, make it into microspheres through method of spray drying. In order to extend its half-life, improve the bioavailability, better to play its curative effect.
Objective:Research the best extraction and purification technology of fructus gardenia antidepressant active site, and prepare it into microspheres.
Methods:With the cumulative swimming fixed time as the assessment index, to optimize active fraction of Gardenia by mice depressed model. The best extraction technology of fructus gardenia was optimized by L_9(3~4) orthogonal experiment with the anointed rate and geniposide content as the assessment index, by the method of HPLC to survey the content of geniposide .To optimized the best purification technology, using the method of macroporous adsorption resin of fructus gardenia coarse extracts of purification, through of macroporous adsorption resin type, adsorption speed, adsorption time, elution concentration, elution speed and volume of use elution as the assessment index. By spray drying microspheres are prepared, the best carrier material is determined by single factor experiment, the effect of drugs than with carrier material, inlet temperature, outlet temperature, feed flow rate on the degree of encapsulated efficiency, yield, the envelope yield are investigated by orthogonal experiment, by which the optimum Preparation Technology is determined. Meanwhile, check the form of microspheres, particle diameter and distribution, burst effect,and by means of measuring the release degrees In vitro,to preliminary study the fructus gardenia antidepressant active site microspheres in Drug delivering mechanism.
Results:60% ethanol extract of fructus gardenia have apparente anti- depression effect. The best extraction technology of fructus gardenia is: add 6 times of 60% ethanol solution, extract three times, each time 1h; The purification technology of geniposide is: the type of macroporous resin is AB-8, the rate of uploading is 2.0mL/min, the amount of uploading is 2BV, the concentration of uploading is 0.25g crude drug /ml of fructus gardenia extracting solution, adsorb 1h, wash with 5BV water for eluting impurity, then used 6BV 40% ethanol as eluent, the rate of elution was 3.0mL/min. the method of prepare microsphere is Spray drying .The best technology is :the ratio of the drugs and the carrier material is 1:, 130℃is intake airflow temperature and extake airflow temperature is 90℃, the rate of sample injection is 300ml/h. the release behavior of geniposide of anti-depression active site in microsphere is agree with higuchi model.
Conclusion:The process is simple and reasonable, and suitable for big industrial production.
目的:研究栀子抗抑郁活性部位的最佳提取、纯化工艺,并将其制备成微球。
方法:通过小鼠抑郁模型,以累计游泳不动时间为考察指标,筛选栀子的抗抑郁活性部位。采用L_9(3~4)正交试验法,以出膏率和栀子苷的含量为考察指标,HPLC法测定栀子苷的含量,优选栀子最佳提取工艺;采用大孔吸附树脂法对栀子的粗提取物进行纯化,通过对大孔吸附树脂的型号、吸附速度、吸附时间、洗脱剂浓度、洗脱速度及洗脱剂用量等因素的考察,确定最佳纯化工艺;采用喷雾干燥法制备微球,通过单因素试验法,确定最佳载体材料;通过正交试验法,考察药物与载体材料比、进风温度、出风温度、进料速度,以收率、包封产率为评价指标,确定微球的最佳制备工艺;同时对微球的形态、粒径及其分布、突释效应进行检查,通过测定其体外释放度,进行栀子抗抑郁活性部位微球释药机理的初步研究。
结果:栀子60%乙醇提取物具有明显的抗抑郁作用;栀子最佳提取工艺为加6倍量60%的乙醇,提取3次,每次1h;栀子苷的纯化工艺为AB-8型大孔吸附树脂,以2ml/min的速度上样2倍柱体积0.25g生药/ml的栀子提取液,吸附1h,先用5倍柱体积的水洗脱杂质,再用6倍柱体积的40%乙醇以3ml/min的速度进行洗脱;采用喷雾干燥法制备微球,最佳工艺:药物与载体材料比为1:2,进风温度为130℃,出风温度为90℃,进料速度为300ml/h;结果显示栀子抗抑郁活性部位微球中栀子苷的释药行为符合Higuchi模型。
结论:工艺简单合理、适合大工业生产。
Fructus gardenia is the dry ripe fruit of Mountain gardenia Rubiaceae, cold-natured, bitter in taste, innocuity, with the function of reducing fire except vexed, clearing away heat evil and promoting diuresis, cooling blood eliminating stasis to activate blood circulation. The drug is one piece of the 2010 edition of China pharmacopoeia, it is mainly used for treat fever upset, acute icteric hepatitis, cystitis, conjunctival congestion score pain, upper gastrointestinal hemorrhage and partly to bleed, bruised. At present the pharmacological research of fructus gardenia is increasing, main is concentrated in resistance to asthma action, anti-inflammatory analgesic action, cholaneresis hepatoprotective effect, antiviral effect and the function of nervous system etc. The early experiment proofed the ethanol extract of fructus gardenia has antidepressant activities. However ,the geniposide which is the active of fructus gardenia the feature of the short of half-life.So this subject adopts ethanol refluxing extraction geniposide, uses the macroporous adsorption resin in purification, And through the pharmacodynamics experiment verify the extract of fructus gardenia antidepressant effect, make it into microspheres through method of spray drying. In order to extend its half-life, improve the bioavailability, better to play its curative effect.
Objective:Research the best extraction and purification technology of fructus gardenia antidepressant active site, and prepare it into microspheres.
Methods:With the cumulative swimming fixed time as the assessment index, to optimize active fraction of Gardenia by mice depressed model. The best extraction technology of fructus gardenia was optimized by L_9(3~4) orthogonal experiment with the anointed rate and geniposide content as the assessment index, by the method of HPLC to survey the content of geniposide .To optimized the best purification technology, using the method of macroporous adsorption resin of fructus gardenia coarse extracts of purification, through of macroporous adsorption resin type, adsorption speed, adsorption time, elution concentration, elution speed and volume of use elution as the assessment index. By spray drying microspheres are prepared, the best carrier material is determined by single factor experiment, the effect of drugs than with carrier material, inlet temperature, outlet temperature, feed flow rate on the degree of encapsulated efficiency, yield, the envelope yield are investigated by orthogonal experiment, by which the optimum Preparation Technology is determined. Meanwhile, check the form of microspheres, particle diameter and distribution, burst effect,and by means of measuring the release degrees In vitro,to preliminary study the fructus gardenia antidepressant active site microspheres in Drug delivering mechanism.
Results:60% ethanol extract of fructus gardenia have apparente anti- depression effect. The best extraction technology of fructus gardenia is: add 6 times of 60% ethanol solution, extract three times, each time 1h; The purification technology of geniposide is: the type of macroporous resin is AB-8, the rate of uploading is 2.0mL/min, the amount of uploading is 2BV, the concentration of uploading is 0.25g crude drug /ml of fructus gardenia extracting solution, adsorb 1h, wash with 5BV water for eluting impurity, then used 6BV 40% ethanol as eluent, the rate of elution was 3.0mL/min. the method of prepare microsphere is Spray drying .The best technology is :the ratio of the drugs and the carrier material is 1:, 130℃is intake airflow temperature and extake airflow temperature is 90℃, the rate of sample injection is 300ml/h. the release behavior of geniposide of anti-depression active site in microsphere is agree with higuchi model.
Conclusion:The process is simple and reasonable, and suitable for big industrial production.
引文
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[2]王伟民,王松龄,任德启.乌灵胶囊为主治疗抑郁症的临床研究[J].辽宁中医药大学学报,2006,8(5):68～69.
[3]李涛.抗抑郁药物的作用机理与研究进展[J].齐鲁药事,2010,29(8):481～483.
[4]王景霞.贯郁胶囊的药学研究及其治疗抑郁症的药效学研究和机理探讨[D].北京中医药大学,2005:1～157.
[5]郝文宇.栀子粗提物治疗慢性应激致抑郁模型小鼠的机制研究[D].北京协和医学院,2009:1～55.
[6]汪玉芳.心达欣胶囊对抑郁模型小鼠行为学的影响及抗抑郁作用机理的研究[D].暨南大学,2009:1～54.
[7]程工倪.KYY01治疗抑郁症的药效学研究和机理探讨[D].北京中医药大学,2009:1～95.
[8]张建忠.国外抗抑郁药的研究进展及其国产化现状[J].上海医药情报研究,2004,1(72):1～3.
[9]韩迎辰.抑郁症及抗抑郁天然药物研究进展[J].药学实践杂志,2005,23(1):3～5.
[10]周学东,陈兴宝.抑郁症经济负担研究进展[J].上海医药,2006,27(12):539～541.
[11]蒋麟.以越鞠丸为基础的抗抑郁中药复方药理作用及其机理研究[D].成都中医药大学,2004:1～55.
[12]何咏梅,杜绍礼.抗抑郁中药的研究进展[J].湖南中医药导报,2003,9(6):71～73.
[13]宋文喜.郁舒颗粒治疗抑郁症的临床与实验研究[D].中国中医研究院,2003:1～56.
[14]国家药典委员会.中华人民共和国药典2010版一部.北京.化学工业出版社.
[15]刘国敏,郭素华,程维明.栀子的药理作用及其机制研究新进展[J].海峡药学,2008,20(11):8～10.
[16]于维萍,傅春升,钟映莉等.栀子中环烯醚萜类化合物的研究概况[J].山东中医杂志,2007,26(8):579～581.
[17]黄仕孙,吴曙粤.栀子的现代药理研究及临床应用概述[J].内科,2010,5(5):534～536.
[18]胡震,杨广德,罗国安等.栀子中栀子苷提取工艺及HPLC分析[J].中成药,2006,28(3):336～338.
[19]吕茂平,乔庆彬,庞春燕等.大孔树脂对栀子苷分离效果的研究[J].中草药,2002,33(9):794～796.
[20]倪慧艳,张朝晖,傅海珍.中药栀子的研究与开发概述[J].中国中药杂志,2006,31(7):538～541.
[21]于洋,高昊,戴毅等.栀子属植物化学成分的研究进展[J].中草药,2010,41(1):148～153.
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