首页 > 硕士 > 医学 > 正文

2-甲氧基雌二醇脂质体冻干粉的研究

The Study on the Lyophilized Liposomes Containing 2-Methoxyestradiol

作者: 专业:药物分析学 导师:杜斌 年度:2010 学位:硕士  院校: 郑州大学

Keywords

2-Methoxyestradiol, liposome, pharmacokinetic, tissue injuries

        2-甲氧基雌二醇(2-methoxyestradiol,2-ME)是雌激素的天然代谢产物,因其具有广谱抗肿瘤活性,且毒性较低,近年来被广泛关注,口服胶囊已被FDA批准完成二期临床研究。然而,2-ME体内胃肠道吸收差,血浆消除速率快,口服或腹腔注射大剂量2-ME才能达到抑制肿瘤的作用。脂质体作为抗癌药物的载体,在某种程度上可提高化疗药物的靶向性和化疗药物的治疗指数,降低用药剂量并大幅降低药物的毒副作用。本研究拟将2-ME制成脂质体冻干粉,以改变体内的释放特性,提高疗效,降低毒副作用,本研究主要内容包括以下几方面:1、对2-ME脂质体的处方和工艺进行考察,以包封率为指标,筛选出注入法为最佳制备方法。通过选择磷脂的种类、有机溶剂的种类、药脂比、冻干保护剂的种类和用量、磁力搅拌时间和超声功率,确定脂质体冻干粉的处方组成和制备工艺,并进行正交试验优化处方,最终制备的2-ME脂质体冻干粉包封率为75%,平均粒径在220nm左右。透射电镜下观察,复溶后脂质体的外观圆整,分布均匀。稳定性实验表明,2-ME脂质体冻干粉4℃保存6个月内稳定性良好。2、建立了以来曲唑为内标,HPLC法测定大鼠血浆样品中2-ME浓度的方法,并对其进行方法学确证。结果显示,该方法线性关系良好,日内和日间精密度均小于11.30%,平均提取回收率为86.2%,生物样品在室温、冷藏、冰冻以及反复冻融情况下稳定性良好,该方法可用于2-ME在大鼠体内血浆中药物浓度的测定。3、SD大鼠的药动学实验:分别按10mg/kg的剂量尾静脉注射2-ME溶液和2-ME脂质体,在设定的时间点眼眶后静脉丛取血,测定血药浓度。用3P97程序求算药动学参数,进行药物动力学模型拟合。结果表明,2-ME在大鼠体内的药动学符合二室模型,同时静脉注射2-ME脂质体和2-ME溶液,2-ME脂质体可以快速到达靶器官,提高治疗效果。4、昆明种小鼠的组织损伤实验表明,2-ME脂质体组对实验动物的毒副作用和组织损伤均明显减轻,组织损伤实验结果验证2-ME脂质体达到降低药物毒性的预期目标。
    2-Methoxyestradiol(2-ME), formerly regarded as an inactive metabolite of estradiol, is now known to be an anti-angiogenic agent with anti-proliferative and cytotoxic activity that confers anti-tumour activity in some animal models of solid tumour.2-ME is currently being evaluated in PhaseⅡclinical trials for the treatment of solid tumors and is undergoing preclinical evaluation for inflammatory conditions. However, poor bioavailability, resultant lack of efficacy and the estrogenic actions of 2-ME show that 2-ME is a suboptimal therapeutic agent in its current formulation. Liposomes are well recognized as drug delivery vehicles that have target effents, delayed drug release and degraded drug toxicity. The purpose of the study is to prepare 2-ME liposome to improve its therapeutic efficacy.The effects of the methods of preparation, the type of phospholipid and organic solvent, the ratio of drug to lipid, the types and amount of lyprotectant, the time of magnetic stirring and ultrasonic power were studied. Then based on the results of single factor analysis, the formulation and the preparation techniques were optimized by orthogonal design.2-ME liposomes were prepared by injection method. The optimized formulation of 2-ME liposomes was as follows:The quality ratio of phosphotidylcholine and cholesterol was 10:1, The quality ratio of lipids and drug was 12:1, The volume ratio of H2O and oil was 3:1, The concentration of poloxamer 188 was 0.20%.To resolve the instability of liposome dispersion, freeze-dried technique was utilized to prepare freeze-dried liposomes. Its main advantage is of high stability because of its solid form during preservation. Mannitol and trehalose were selected as the optimum cryoprotectants due to the minimum change of the average particle sizes and the entrapment efficiency before and after freeze-drying process. The size of freeze-dried liposomes after hydration was about 220nm. The entrapment efficiency was about 75%. The transmission electron microscopy showed that most liposomes were spherical. The stability test was conducted, the results showed that 2-ME freeze-dried liposomes kept at 4℃for 6 months showed good stability.The HPLC method based on fluorescence detection for the quantitative determination of 2-ME in plasma samples using letrozole as the internal standard, has been developed and validated. There was good linear relationship for 2-ME with the ranges in plasma samples. The intra-day and inter-day precision of quality control samples were less than 11.30%. The average recovery of extraction was 86.2%. Biological samples were stable in room temperature or 4℃, and in freezing or after three freeze/thaw cycles.Pharmacokinetic parameter calculation and pharmacokinetic model were carried out using the 3P97 pharmacokinetics software. In vivo pharmacokinetic studies data indicated that the 2-ME solution followed a two-compartment model while 2-ME liposome suspension also followed two-compartment model with different pharmacokinetic parameters after i.v. administration of liposomes and solution at a dose of 10 mg/kg to rats. The liposomes have good biocompatibility and biodegradability, so it might promote its distribution in tissues, the drug could rapidly act on therapeutic effect.Kunming mice acute toxicity experiments showed that 2-ME liposomes significantly reduced toxicity and tissue injury compared to 2-ME solution. The results verified that 2-ME liposome was security.
        

2-甲氧基雌二醇脂质体冻干粉的研究

摘要4-5
Abstract5-6
目录7-12
前言12-14
第1章 2-甲氧基雌二醇及脂质体冻干粉的研究进展14-25
    1 2-ME的研究进展14-17
        1.1 抗肿瘤作用机制14-15
        1.2 2-ME作为抗癌药物的优势15-16
        1.3 小结16-17
    2 脂质体冻干粉的研究进展17-25
        2.1 冻干机制17-18
        2.2 残留水量的控制18-19
        2.3 T_m,T_g和ESR19
        2.4 处方因素19-22
        2.5 工艺参数22-23
        2.6 结论23-25
第2章 2-甲氧基雌二醇脂质体处方前研究25-34
    1 仪器与试剂25-26
        1.1 试剂25
        1.2 仪器25-26
    2 方法26-28
        2.1 2-ME理化性质的考察26-27
        2.2 2-ME含量测定方法的建立27-28
        2.3 2一甲氧基雌二醇脂质体包封率的测定28
    3 结果28-32
        3.1 2-ME检测波长的确定28-29
        3.2 2-ME的酸碱性29
        3.3 紫外分光光度法测定2-ME的解离常数29
        3.4 2-ME水中溶解度的测定29-30
        3.5 2-ME含量测定方法的建立30-31
        3.6 2-ME脂质体包封率的测定31-32
    4 讨论32-33
        4.1 色谱条件的确定32
        4.2 包封率的测定32-33
    5 本章小结33-34
第3章 2-甲氧基雌二醇脂质体的制备34-43
    1 仪器与试剂34-35
        1.1 试剂34
        1.2 仪器34-35
    2 方法35-37
        2.1 脂质体制备方法筛选35-36
        2.2 处方筛选36-37
        2.3 工艺优化37
        2.4 包封率的验证37
    3 结果37-41
        3.1 不同制备方法结果37-38
        3.2 有机溶剂的选择38
        3.3 磷脂种类的选择38-39
        3.4 正交试验结果39-40
        3.5 工艺优化40-41
        3.6 包封率的验证41
    4 讨论41-42
        4.1 制备方法的选择41
        4.2 药脂比对包封率的影响41-42
        4.3 胆固醇用量的影响42
        4.4 制备工艺的影响42
    5 本章小结42-43
第4章 2-ME-L冻干粉的制备及其质量评价43-54
    1 仪器与试剂43-44
        1.1 试剂43
        1.2 仪器43-44
    2 方法44-46
        2.1 2-ME脂质体冻干粉的制备44
        2.2 粒径及zeta电位的测量44
        2.3 形态学分析44-45
        2.4 粘度的测定45
        2.5 初步稳定性研究45
        2.6 体外释药行为的研究45-46
    3 结果46-52
        3.1 单用冻干保护剂的筛选46-47
        3.2 冻干保护剂的联合使用47-48
        3.3 粒径及zeta电位的测量48-49
        3.4 形态学分析49
        3.5 粘度的测定49-50
        3.6 初步稳定性研究50-52
        3.7 体外释药行为52
    4 讨论52-53
        4.1 冻干保护剂的筛选52-53
        4.2 脂质体表征53
        4.3 稳定性测定53
    5 本章小结53-54
第5章 2-ME-L冻干粉药代动力学研究54-63
    1 仪器与试剂54-55
        1.1 试剂54
        1.2 仪器54
        1.3 实验动物54-55
    2 方法55-57
        2.1 色谱条件55
        2.2 溶液配制55-56
        2.3 样品的处理56
        2.4 方法学考察56-57
        2.5 药动学研究57
    3 结果57-61
        3.1 专属性57-58
        3.2 标准曲线58-59
        3.3 精密度59
        3.4 回收率59
        3.5 稳定性实验59-60
        3.6 大鼠血浆中药物浓度的测定60
        3.7 血浆药动学房室模型的判断60-61
        3.8 房室模型的药动学参数研究61
    4 讨论61-62
        4.1 提取溶剂的选择61-62
        4.2 给药溶剂的选择62
        4.3 检测器选择和内标的选择62
        4.4 药动学参数62
    5 本章小结62-63
第6章 2-ME-L冻干粉组织损伤实验63-68
    1 仪器与试剂63
        1.1 试剂63
        1.2 仪器63
        1.3 实验动物63
    2 方法63-64
        2.1 饲养方法63-64
        2.2 剂量与分组64
        2.3 观察期64
    3 结果64-67
        3.1 笼旁观察与系统尸解64
        3.2 病理切片64-67
    4 讨论67
        4.1 笼旁观察67
        4.2 病理切片67
    5 本章小结67-68
结论68-69
参考文献69-76
致谢76-77
个人简历77
        下载全文需77


本文地址:

上一篇:冠心Ⅱ号不同配伍方及不同剂型对丹参素、阿魏酸药代动力学的影响
下一篇:最后一页

分享到: 分享2-甲氧基雌二醇脂质体冻干粉的研究到腾讯微博           收藏
评论排行
公告