注射用洋参生脉粉针的研究

【作者】 尹湉；

【导师】 唐星；

【作者基本信息】 沈阳药科大学 ， 药剂学， 2006， 硕士

【摘要】 本课题参照部颁标准中生脉注射液的处方及诸多相关文献，将西洋参、五味子、麦冬配伍，对三味中药分别进行提取及纯化精制，制备成注射级中间体干燥品；分别对其进行质量研究并建立相应专属性质量标准；以制得的中间体为原料，筛选出合适的处方和冷冻干燥工艺，制备注射用洋参生脉粉针；制定准确、严密的质量标准并建立麦冬、五味子药材及其中间体和制剂的指纹图谱，从而有效控制产品质量；考察注射用洋参生脉的一般安全性，以保证临床用药的安全；对本品在改善心肌缺血及心律失常方面进行初步药效研究，为临床给药奠定基础。首先，针对注射剂要求及工业化生产规模，采用单因素考察及正交设计试验，以各自有效成分的转移率为指标，对处方中三味中药分别进行乙醇提取及大孔树脂纯化工艺的筛选与优化，确定了各注射级中间体的制备工艺。以西洋参二醇组皂苷（Rd、Rc、Rb₃、Rb₂、Rb₁、Rg₃）及pF₁₁的转移率为指标，制备注射用西洋参二醇组皂苷，制备过程中其总转移率为64.9％；以甾体总皂苷（以麦冬皂苷D′计）、总黄酮（以橙皮苷计）的转移率为指标，制备注射用麦冬标准化提取物，制备过程中其总转移率分别为64.3％、66.4％；以总木脂素（五味子醇甲、五味子甲素及五味子乙素三种有效成分之和）的转移率为指标，制备注射用五味子标准化提取物，制备过程中其总转移率为65.7％。对三个注射级中间体干燥品进行质量研究并建立相应专属性质量标准。检测结果显示三者均符合注射级质量要求，可直接作为二次配料制备注射剂。运用HPLC-ELSD法对注射用西洋参二醇组皂苷中的各单体皂苷同时进行含量测定，规定西洋参二醇组皂苷（Rd、Rc、Rb₃、Rb₂、Rb₁、Rg₃）及pF₁₁的含量总计不低于70％；并测定了总皂苷含量，规定其总皂苷含量不低于95.0％。测定了注射用麦冬标准化提取物中总皂苷及总黄酮含量，规定二者含量分别不低于60.0％、5.0％。采用HPLC法，对注射用五味子标准化提取物中的五味子醇甲、五味子甲素、五味子乙素进行含量测定，规定三者含量总计不低于95.0％；同时测定了总木脂素含量，规定其总木脂素含量不低于58.0％。以外观、含水量和复溶性为指标，优化冻干制剂处方，确立了甘露醇100mg／瓶作为支撑剂的处方及冻干工艺。配伍试验研究表明本品和生理盐水注射液，5.0％葡萄糖注射液配伍相容性良好（12h）。对注射用洋参生脉粉针进行质量研究，结果表明产品符合中药注射用无菌制品的质量要求，并建立专属性质量标准，以有效控制产品质量，保证临床用药的安全。分别运用HPLC-ELSD法测定各西洋参单体皂苷的含量、HPLC法测定五味子醇甲的含量、紫外-分光分光光度法测定总皂苷、总木脂素的含量：规定本品每瓶含西洋参二醇组皂苷（Rd、Rc、Rb₃、Rb₂、Rb₁、Rg₃）及pF₁₁总计不低于21.0mg；五味子醇甲不低于0.38mg；总皂苷（以人参皂苷Rd计）不低于67mg；含总木脂素（以五味子醇甲计）不低于1.50mg。建立了麦冬、五味子药材及其中间体和制剂的指纹图谱。结果表明，药材及其中间体和制剂中指纹图谱方法学考察符合要求，10批药材及其中间体和制剂间相对保留时间、非共有峰峰面积、共有指纹峰峰面积比值等各项指标均在规定范围以内，药材及其中间体和制剂间指纹图谱相关性良好。对注射用洋参生脉粉针的体外溶血、过敏性和血管刺激性进行了考察，结果表明本品的安全性达到注射剂的要求。小鼠急性毒性试验的LD₅₀及95％可信限为492.9mg／kg和438.2～554.5mg／kg（以总皂苷与总木脂素总量计）。药效学试验结果表明，本品大鼠腹腔给药16.8mg／kg以上，对Iso所致大鼠心电图变化有明显的抑制作用；静脉给药24mg／kg以上，对夹闭小鼠气管其心电消失时间有显著的延长作用、对氯仿诱发小鼠室颤的发生率有明显的抑制作用；静脉给药16.8mg／kg以上对氯化钡引起的心律不齐维持时间有显著缩短作用。提示注射用洋参生脉粉针可用于缺血性心脏病及心律失常的治疗。更多还原

【Abstract】 This thesis aimed to investigate the extractions of Panax quinquefolium L., Ophiopogon japonicus and Schisandra Chinensis and the combination of three extracts. The freezing-dry intermediates for injection of three extracts were manufactured and quality specification were established. To prepare Yangsheng Shengmai Freezing-dry Preparation （YSFP） for injection from the intermediates and maintain the qualities of the product, the formula and the and drying processes were optimized; an accurate and sensitive quality specification of freezing-dry products and the fingerprint chromatograms of Ophiopogon japonicus, Schisandra Chinensis, the intermediates and the preparation were established. The investigation of common safety and pharmacodynamics experiments provided experimental data for clinical application of YSFP.The extracting with alcohol and purifying with macroporous resin processes of three kinds of Chinese traditional medicine were optimized by using factor experiment and orthogonal design, As the response, the transfer proportion of each active ingredient was evaluated. The formulation and preparation process of each intermediates for injection were determined according to result of experiments.The purification process of the active constituents of Panax quinquefolium L. is studied by the criterion of transference rate of Rd, Rc, Rb₃, Rb₂, Rb₁, Rg₃, pF₁₁. The result indicated that the total transference rate was 64.9%. The purification process of the active constituents of Ophiopogon japonicus is studied by the criterion of transference rate of total saponins and total flavonoids. The result indicated that the total transference rate was 64.3%, 66.4%. The purification process of the active constituents of Schisandra Chinensis is studied by the criterion of transference rate of total lignans. The result indicated that the total transference rate was 65.7%.Qualities of the three intermediates for injection were studied separately and built up corresponded. The results showed that all of them were agree with the quality requirements of preparation for injection.HPLC-ELSD method was used to determine the sing saponin from PQDS, which regulated all of the contents of Rd, Rc, Rb₃, Rb₂, Rb₁, Rg₃, pF₁₁ should be more than 70.0%, at the same time. The contents determination on total saponins were determined and regulated more than 60.0%. The contents determination for Ophiopogon japonicus intermediate on total saponins and total flavonoids were determined and regulated more than 60.0% and 5.0%. HPLC and UV methods were set up to determine contents of lignans in the Schisandra Chinensis intermediate and regulated more than 15.0%, 58.0%.The prescription of freezing-dry preparation was optimized by the criterion of appearance, water content and re-solubility. Mannitol （100mg/bottle） was used as filler.The quality of YSFP was studied, and the quality specification was built up to agree with the quality requirements of Chinese herbal freeze-drying preparation for injection. The quality could be controlled by the specification established considerably and the clinical safety could be ensured.HPLC-ELSD method for sing saponins, HPLC method for schizandrol A, UV methods for total saponins and total lignans were used to determine their contents in the preparation, which regulated the determination should be more than 21.0, 0.38, 67.0, 1.50mg/bottle.Fingerprint chromatograms of Ophiopogon japonicus, Schisandra chinensis, the intermediates and the preparation were established to control their qualities. Results of tests for HPLC methods of the fingerprint chromatograms were up to grade. Total area ratio of all the uncommon peaks, relative retention time and relative area ratio of the common peaks all met the related requirements for fingerprint chromatograms of 10 batches of Ophiopogon japonicus, Schisandra chinensis, the intermediates and the preparation.To assess whether there was a potential risk of hemolysis, stimulus and allergy, experiments were proceed and the result showed that the product was safety. The acute toxicity test of YSFP concluded that the LD₅₀ and confidential interval （95%）, correspondingly, were 492.9mg/kg and 438.2～554.5 mg/kg.The result of pharmacodynamics studies showed that there was significant inhibition on changes of electrocardiogram caused by isoprenaline hydrochloride injection after intraperitoneal injection of YSFP to rats（16.8mg/kg）; significant prolongation of electrocardio disappear time and decrease on incidence rate of cardiac ventricle jitter caused by chloroform after intravenous injection of YSFP to mice（24mg/kg）, significant decrease of maintaining time of arrhythmia caused by barium chloride after intravenous injection of YSFP to mice（16.8mg/kg）. All these data indicated its potential application on therapy of ischemic heart disease and arrhythmia.更多还原