【作者】 张丽娜；

【导师】 张振中； 张正全；

【作者基本信息】 郑州大学 ， 药物分析学， 2009， 硕士

【摘要】 2-甲氧基雌二醇（2-methoxyestradiol,2-ME）是雌激素的天然代谢产物,具有抗肿瘤活性,目前,其抗肿瘤活性国外处于Ⅰ、Ⅱ期临床研究阶段,2-ME是一种浓度时间依赖性的抗肿瘤药物,在水中溶解度极低(约0.2206μg·ml^-1),口服吸收和剂量之间缺乏相关性,在体内又易快速羟基化、脱氢等生物变化产生非活性的代谢产物,导致半衰期较短,生物利用度较低。因而考虑将其制成乳剂以达到缓释及提高生物利用度的作用。本研究的目的是以2-ME为模型药物制备静脉注射乳剂,考察脂溶性药物制成O/W乳剂后的安全性,药效性及缓释靶向性,为2-ME的新剂型开发提供研究基础。本文对2-ME静脉乳剂的处方和工艺进行了研究,测定了25℃时,2-ME在大豆油、水中的溶解度分别为1.05 mg·ml^-1、0.2206μg·ml^-1,脂水分配系数logP为4.41。考察了乳剂处方中油相比例,乳化剂（蛋黄卵磷脂、泊洛沙姆）比例,药物与磷脂比例及乳化温度、乳化时间、均质压力及均质次数等影响乳化效果的处方及制备工艺因素,用正交设计及单因素法筛选出较优的处方及制备工艺。对2-ME静脉注射乳剂药剂学性质进行了研究,测得2-ME乳剂的粒径为246.2±5.6nm;Zeta电位为-32.47±1.51 mv;pH值为7.32±0.12;黏度为1.423±0.005 mpa.S,符合2005版药典对乳剂的要求。通过离心稳定性,影响因素试验,冷冻-加热循环试验,加速试验及长期试验的研究,表明本研究所制备的乳剂稳定性较好,在冷藏条件下粒径无明显变化,有较好的贮存稳定性。通过不同的释药条件及释药方法对2-ME乳剂在体外的释放情况进行考察,2-ME对照液有突释现象,30 min达50%,2 h达到90%,以后缓慢释放,2-ME乳剂的累积释药量则在6 h才接近50%,30 h接近90%,可见在体外2-ME乳剂有一定的缓释效果。通过对2-ME乳剂进行急性毒性、刺激性、过敏性以及溶血性检查,对其安全性进行初步评价。结果显示2-ME静脉注射乳剂毒性非常低,对兔耳静脉血管没有不良刺激作用,无溶血及过敏反应发生。采用MTT体外药敏实验,验证了2-ME乳剂对MCF-7细胞的增殖抑制作用,并且这种抑制作用随着时间及剂量的增加而增强。以小鼠为模型动物,采用HPLC（荧光检测器）测定2-ME在血浆及各组织中的浓度,2-ME静脉注射乳剂在小鼠体内的药物代谢动力学规律;结果表明2-ME静脉注射乳剂具有一定的缓释及肺、肝、脾等组织靶向性。更多还原

【Abstract】 2-Methoxyestradiol（2-ME）,which is studied at clinical phaseⅠ,Ⅱabroad,is a natural estrogen metabolite with the characteristics anti-tumors activity.2-ME has the manners of time-concentration-dependent,low solubility(approximately 0.2206μg·ml^-1);lack of correlation between oral absorption and dose;and it can rapidly produce inactive metabolites due to easy hydroxylation,dehydrogenation in vivo.The above causes resulted in shorter half-life and low oral bioavailability.The emulsion has the roles of delayed-release and increase bioavailability.The purpose of the study is to prepare o/w intravenous emulsion of 2-ME and investigate the safety,efficacy, delayed-release and targeted effect to tumors cells,and provide the research base for new formulations of 2-ME.Formulation design and preparation technique of 2-ME intravenous emulsion was studied,the solubility of 2-ME in soybean oil and water were 1.05 mg·ml^-1,0.2206μg·ml^-1 at 25℃respestively,the lipid water partition coefficient（logP） was 4.41 at 25℃,so 2-ME could be dissolved in soybean oil to prepare emulsion.The effects of the percentage of soya bean oil,the percentage of emulsifier,the ratio of between phospholipids and poloxamer,the ratio of between drug and phospholipid and the influennces of emulsification temperatures,emulsification time,homogeneous stress and homogeneous times on the emulsion quality were studied,Then based on the results of single factor analysis,the formulation and the preparation techniques were optimized by orthogonal design.The pharmaceutical properties of 2-ME intravenous emulsion were as followings: particle size 246.2±5.6 nm,Zeta potential -32.47±1.51 mv,pH value 7.32±0.12, viscosity 1.423±0.005 mpa.S.The physicochemical properties of 2-ME intravenous emulsion were coincident with the Chinese Pharmacopoeia.The centrifugal test, impact factor test,frozen-heating cycle test,accelerated test and long-term test were conducted to investigate the stability of emulsion,the results showed that the particle size didn’t increase significantly,indicating that the emulsion had good stability in frozen conditions.The accumulated release amount of 2-ME control solution was up to 50%in 30 min,up to 90%in 2 h,the accumulated release of 2-ME emulsion was about 50%in 6 h,about 90%in 30 h.Showing that 2-ME emulsion had slow-release effect compared to 2-ME control solution.The preliminary safety of 2-ME emulsion was evaluate by acute toxicity test, vascular irritation test,haematolysis and hypersensitiveness test.The results showed that the emulsion had no obvious toxicity,irritation,haemocytolysis and hypersensitiveness.The effects of the 2-ME control solution and 2-ME emulsion on the MCF-7 cell proliferation inhibition were investigated by use of MTT method, indicating that inhibition effect was increased with time and dose increment, 2-ME-emulsion had slow-release effect compared to 2 -ME control solution.2-ME blood concentration of mice were determined by HPLC with fluorescence detector to explore pharmacokinetics parameters for 2-ME intravenous emulsion and 2-ME control;At the same time,the 2-ME distribution in mice were investigated for 2-ME intravenous emulsion.Showing that 2-ME emulsion has a sustained-release and tissue targeting in lung,liver,spleen compared to 2-ME control solution.更多还原