2-甲氧基雌二醇注射混悬剂的研究

发布时间：2018-02-28 21:00

本文关键词： 2-甲氧基雌二醇混悬剂药代动力学药效学体内外相关性　出处：《郑州大学》2014年硕士论文　论文类型：学位论文

【摘要】：2-甲氧基雌二醇（2-Methoxyestradiol，简称2-ME）是一种雌激素的天然代谢物，人体内非常少。近年来国内外研究发现，该化合物对多种肿瘤细胞都有较好的抑制作用，且具有对正常细胞毒性低、机体耐受性良好的特点。目前国外将2-ME作为一种比较新型的抗肿瘤药物进行研究，现在处于Ⅱ期临床研究阶段。然而，2-ME是一种难溶于水的药物，在水中的溶解度非常低，口服存在肝首过效应，生物利用度低等问题；其注射剂体内代谢非常快，半衰期短，在体内滞留时间短，要长期维持有效的血药浓度变得很难。因此，如何提高并维持2-ME在体内的有效血药浓度是其制剂学研究的主要问题。本文以2-ME为主药，添加不同的辅料制备2-ME注射混悬剂，通过肌肉注射延长释放时间，达到缓释作用，减少给药次数，，提高病人的依从性。本文首先建立了高效液相色谱法用于测定2-ME的含量和体外释放的研究，流动相为甲醇:水(70:30)，流速为1.0mL·min-1，柱温30℃，紫外检测器（288nm）。以药物浓度和峰面积进行线性回归得到2-ME的回归方程：2-ME的定量方程为y=0.2949x+0.0752(R2=0.9999)，日内、日间精密度分别为1.33%、3.45%，平均回收率为99.97%（RSD=1.32%）。该方法符合样品分析的要求。并用高效液相色谱法测得2-ME原料药在37℃水中的溶解度为（1.579±1.131）μg·mL-1，2-ME的3襊脂水分配系数为（2.79±1.43）。本文通过单因素试验分别考察了润湿剂、助悬剂、表面活性剂对混悬剂的重新分散性，稳定性和沉降体积比的影响，并通过正交试验确定了药物和各种辅料的用量，最终选用2-ME，CMC-Na及泊洛沙姆188的含量分别为70，8，6mg·mL-1。并采用分散法制备2-ME注射混悬剂。然后对2-ME注射混悬剂进行了评价。实验结果显示：2-ME注射混悬剂为乳白色的溶液，沉降体积比为98%、重新分散性、通针性良好、平均粒径8.31微米，在温度、湿度、光照等条件下也很稳定，各种质量标准均符合药典对于混悬剂的规定。同时本论文考察了2-ME注射混悬剂的体外释放，释放介质为0.3%SDS溶液，释放温度为37℃，选择流通池滴流法考察制剂的体外释放。通过对2-ME混悬剂中药物的体外释放模型进行拟合，结果表明该制剂符合一级级释放模型，36h2-ME释放了92.69%基本释放完全。本文通过小鼠肌肉注射给药后，取血浆后经处理考察2-ME混悬剂的药动学特点，结果混悬剂在小鼠体内成单室模型，半衰期（t1/2Ke）为（7.52±0.26）h，平均体内滞留时间（MRT）为（16.18±1.25）h，AUC0→48h为（27.81±2.09）μg·mL-1·h。同时通过测量肌肉中剩余药量考察制剂的体内释放，实验结果显示，2-ME在48h释放了82.82%基本完全释放。体内释放的释放特性用Ritger-Peppas指数模型拟合，药物以扩散机理释放，且药物释放方式为Fick扩散。并通过点对点相关对2-ME混悬剂的体内外释放进行了相关性分析，相关系数r=0.9909，说明体内外释放数据具有一定的相关性。本研究采用皮下接种肿瘤细胞悬液的方法建立S180腹水瘤模型，以小鼠体重、肿瘤体积变化、相对肿瘤增殖率、抑瘤率为考察指标，初步研究了2-ME注射混悬剂的体内抗肿瘤效果。结果显示2-ME注射混悬剂相对肿瘤增殖率为31.51%~63.84%，抑瘤率大于60%，对小鼠腹水瘤有抑制作用，但未达到预期效果，需要进一步的研究。
[Abstract]:2- methoxyestradiol (2-Methoxyestradiol, referred to as 2-ME) is a natural metabolite of estrogen, the body is very small. Recent studies have shown that this compound has good inhibitory effect on tumor cells, and has low toxicity to normal cells, the characteristics of the body well tolerated. At present the foreign 2-ME as a new anticancer drug research, now in phase II clinical trials. However, 2-ME is a drug which is difficult to dissolve in water, very low solubility in water, oral hepatic first pass effect, low degree of biological utilization; the injection in vivo metabolism is very fast, short half-life, in in short retention time, to maintain effective blood concentration becomes very difficult. Therefore, how to improve and maintain 2-ME is the main problem in the research and preparation of effective blood concentration in vivo. The 2-ME based medicine, 2-ME injecting suspensions were prepared by adding different excipients, and the release time was prolonged by intramuscular injection to achieve sustained release, reducing the number of drug delivery and improving the compliance of the patients.
This paper established a HPLC method for determination of content and in vitro release of 2-ME, the mobile phase of methanol water (70:30), the flow rate was 1.0mL - min-1, the column temperature of 30 DEG C, UV detector (288nm). The drug concentration and peak area of linear regression, 2-ME regression equation: quantitative the 2-ME equation is y=0.2949x+0.0752 (R2=0.9999), within day precision were 1.33%, 3.45%, the average recovery rate was 99.97% (RSD=1.32%). This method meets the requirements of sample analysis. The solubility and high performance liquid chromatographic method for measuring the 2-ME API in the water of 37 DEG C (1.579 + 1.131). G mL-1,2-ME 3 Cuo lipid water partition coefficient was (2.79 + 1.43).
In this paper, through the single factor experiment investigated the wetting agent, suspending agent, surface active agent on suspension re dispersion, stability and effect of sedimentation volume ratio, and determine the amount of drug and excipients by orthogonal test, the final selection of 2-ME, CMC-Na and poloxamer 188 Tim content were 70,8,6mg. ML-1. and 2-ME were prepared by dispersion injection suspension.
Then the injection of 2-ME suspension was evaluated. The experimental results showed that injection of 2-ME suspension was milky white, sedimentation volume ratio is 98%, re dispersibility, good syringeability, the average particle size of 8.31 microns, the temperature, humidity, light and other conditions are very stable, quality standards the results met the Pharmacopoeia provisions for suspension of release. At the same time this paper examines the injection of 2-ME suspension in vitro release medium for 0.3%SDS solution, the release temperature is 37 DEG C, select the flow cell drip method was used to study the release of 2-ME. By fitting the suspension in the in vitro drug release model results. The results indicate that the agent meets the first grade level release model, 36h2-ME released 92.69% totally released.
The mice were injected after the pharmacokinetic characteristics of plasma obtained after treatment of 2-ME suspension drug, results suspension into a single compartment model in vivo half-life (t1/2Ke) for (7.52 + 0.26) h, the average body retention time (MRT) for (16.18 + 1.25) H. AUC0, 48h (27.81 + 2.09) g - mL-1 - H. at the same time by measuring the residual amounts of muscle preparation was investigated in vivo release, the experimental results show that 2-ME in 48h has released 82.82% basic complete release. In vivo release release characteristics using Ritger-Peppas index model, the diffusion mechanism of drug release, and drug release for the Fick diffusion. And through the point of 2-ME suspension in vitro and in vivo release for the correlation analysis, the correlation coefficient r=0.9909, illustrate the in vivo release data have a certain correlation.
Based on the method of subcutaneous tumor cell suspension of S180 ascites tumor model, the mice weight, tumor volume changes, the relative tumor proliferation rate, tumor inhibition rate as the study index, preliminary study on antitumor effect of 2-ME suspension for injection in vivo. The results showed that 2-ME injection suspension for 31.51%~63.84% relative tumor growth rate and the inhibition rate more than 60%, has inhibitory effect on mouse ascitic tumor, but did not reach the expected effect, the need for further research.

【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】：R944

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