2-甲氧基雌二醇干粉吸入剂的研究

发布时间：2018-05-13 12:13

本文选题：2-甲氧基雌二醇 + 2-ME可吸入微粒　；参考：《郑州大学》2014年硕士论文

【摘要】：2-甲氧基雌二醇(2-Methoxyestradiol,2-ME)是雌二醇的内源性代谢产物，药理活性研究表明其对多种癌细胞都有较好的作用效果，具有广谱抗肿瘤作用。本实验室之前对其口服制剂和注射剂进行了广泛研究，发现其口服吸收无规律，生物利用度低，注射剂在体内代谢快，半衰期短，不能长时间维持有效的血药浓度。因此积极探索其他的给药方式，改变上述剂型所存在的缺点是极其必要的。干粉吸入剂是通过肺部给药的制剂，其可避免肝脏的首过效应，提高药物的生物利用度，药物在肺部的吸收起效亦较快，且能实现肺部靶向给药，更有利于肺部疾病的治疗。因此，本实验拟将2-ME制成干粉吸入剂，探讨其用于肺部给药的可行性，为肺部肿瘤的治疗开辟新剂型。本文首先建立了2-ME干粉吸入剂的体外HPLC分析方法，经方法学考察知，该分析方法的专属性强，日内、日间精密度分别为0.99%、1.06%，平均回收率为100.18%，均符合应用HPLC分析法的规定，2-ME在0.2～40μg·ml-1的浓度范围内，浓度与峰面积的线性关系良好，所得回归方程及相关系数分别为A=6×107C+106，r=0.9999，因此该方法适用于2-ME体外含量的测定分析。其次，鉴于肺部可吸入微粒粒径的最佳条件为1～5μm范围内，本实验采用控制结晶法先制备了2-ME的可吸入微粒。通过对所制备微粒的外观形态、粒径大小及稳定性的考察，筛选出2-ME微粒制备的最佳处方和工艺条件，分别为：2-ME为2.5mg·ml-1，泊洛沙姆为5mg·ml-1，转速为1200r·min-1，无水乙醇与水的比例为1:3。用上述方法制备出2-ME微粒溶液后，再通过离心、再分散和冷冻干燥制备出2-ME可吸入微粒。第三，以溶解性、粉末形态、引湿性、粒径及其分布、水分含量、休止角、雾化特性、排空率及体外沉积性等粉体理化性质及雾化特性为评价指标，对2-ME干粉吸入剂制备的处方和工艺条件进行研究，得到了2-ME干粉吸入剂制备的最优处方和最佳制备工艺，分别为：①处方，2-ME与乳糖和亮氨酸的比为2:1:1，泊洛沙姆用量为其余三者用量的2%，然后将药物和各辅料配制成一定浓度的混悬溶液，进行喷雾干燥；②制备工艺，进口温度140℃，雾化进气量为470L·h-1，抽真空度为80%，供液速度为240ml·h-1，溶液浓度为1%。第四，，测定了按最优条件制备的干粉吸入剂的各项评价指标，由实验结果知，本品载体溶解性好，能迅速释放出2-ME可吸入微粒；空气动力学径为2.72μm，适合肺部吸入给药；有引湿性，水分含量为2.224±0.73%；外观形态不规则，休止角为42.36°±1.42°，流动性稍差；雾化特性为B级，排空率90%，有效部位沉积量30%，均符合吸入给药的要求。第五，通过建立动物吸入给药模型，对2-ME干粉吸入剂在大鼠体内的初步药代动力学和释放行为进行了考察。由实验结果知2-ME干粉吸入剂在大鼠体内的代谢为一室模型，分布半衰期(t1/2ka)为0.6±0.22h，消除半衰期(t1/2ke)为3.49±1.26h，AUC0-12h为2.95±1.24μg·ml-1·h，释放行为符合Higuchi释放模型，r=0.9945。最后，对2-ME干粉吸入剂肺部给药的安全性进行了初步评价，知2-ME干粉吸入剂对肺部具有一定的刺激性，但无严重刺激性及成纤维细胞增生现象，表明2-ME干粉吸入剂的生物相容性和安全性能够满足肺部给药的要求。
[Abstract]:2- methoxy estradiol (2-Methoxyestradiol, 2-ME) is an endogenous metabolite of estradiol. The pharmacological activity studies show that it has a good effect on various cancer cells and has broad spectrum antitumor effect. Before this laboratory, the oral preparation and injection were extensively studied, and its oral absorption was irregular and bioavailability was found. Low, the injection is fast in metabolism in the body, short half life and can not maintain effective blood concentration for a long time. Therefore, it is extremely necessary to actively explore other ways of administration and change the shortcomings of the above dosage forms. Dry powder inhalant is a preparation through the lung, which can avoid the first over effect of the liver, improve the bioavailability of the drug, and improve the bioavailability of the drug. The absorption of the substance in the lungs is also faster, and it can achieve the lung target drug delivery, which is more conducive to the treatment of lung disease. Therefore, this experiment is to make 2-ME into dry powder inhalant, to explore the feasibility of its use in lung administration, and to open up a new dosage form for the treatment of lung tumor.
In this paper, the method of HPLC analysis in vitro for 2-ME dry powder inhalant is first established. It is known by the jurisprudence that the analysis method has a strong specificity. The day interval is 0.99%, 1.06%, and the average recovery is 100.18%. It is in line with the application of HPLC analysis. 2-ME is in the concentration range of 0.2 ~ 40 mu g. The concentration and peak area are linear. The regression equation and correlation coefficient were A=6 * 107C+106 and r=0.9999 respectively, so the method is suitable for the determination and analysis of 2-ME in vitro.
Secondly, in view of the optimum condition of the inhalable particle size of the lung to be within the range of 1~5 m, this experiment uses the controlled crystallization method to prepare the inhalable particles of 2-ME first. Through the investigation of the appearance, size and stability of the prepared particles, the best formulation and technological conditions for the preparation of 2-ME particles are selected: 2-ME is 2.5mg. Ml, respectively. -1, mooring Losham is 5mg ml-1, the rotational speed is 1200R. Min-1, and the proportion of ethanol and water is 1:3.. The 2-ME particle solution is prepared by the method above, and then the 2-ME inhalable particles are prepared by centrifugation, re dispersion and freeze drying.
Third, on the basis of solubility, powder morphology, wettability, particle size and distribution, moisture content, repose angle, atomization characteristics, the physicochemical properties and atomization characteristics of powders, such as emptying rate and extracorporeal deposition, the formulation and technological conditions of the preparation of 2-ME dry powder inhalers were studied, and the best prescription and the best preparation for the preparation of 2-ME dry powder inhalers were obtained. The best preparation techniques are as follows: (1) the ratio of 2-ME to lactose and leucine is 2:1:1, and the dosage of poloxamer is 2% of the remaining three. Then the drug and the auxiliary materials are prepared into a certain concentration of suspension solution and spray drying; the preparation process is 140, the inlet temperature is 470L. H-1 and the vacuum degree is 80%. The degree is 240ml. H-1, and the solution concentration is 1%.
Fourth, the evaluation indexes of dry powder inhalers prepared on the optimal conditions were measured. The results were known by the experimental results that the carrier was good in solubility and could release 2-ME inhalable particles quickly; the aerodynamic diameter was 2.72 m, suitable for lung inhalation; it was wet, the moisture content was 2.224 + 0.73%, the appearance was irregular and the rest angle was 42.36 The degree of fluidity is slightly worse than the 1.42 degree, the atomization characteristic is B grade, the emptying rate is 90%, and the effective part deposition amount is 30%, which all accord with the requirement of inhalation.
Fifth, the preliminary pharmacokinetics and release behavior of 2-ME dry powder inhalants in rats were investigated by establishing an animal inhalation drug delivery model. The experimental results showed that the metabolism of 2-ME dry powder inhalants in rats was a one chamber model, the distributed half life (t1/2ka) was 0.6 + 0.22h, and the elimination half life (t1/2ke) was 3.49 + 1.26h, AUC0-12h was 2.95 + 1.24 g ml-1 h, the release behavior accords with Higuchi release model, r=0.9945.
Finally, the safety of 2-ME dry powder inhalers was evaluated preliminarily. It was known that 2-ME dry powder inhalers have a certain irritation to the lungs, but there is no serious irritation and fibroblast proliferation. It shows that the biocompatibility and safety of 2-ME dry powder inhalant can be full of the requirements of lung administration.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943

【参考文献】