脂质、P-gp抑制剂及自微乳化释药系统对紫杉醇肠淋巴转运的影响研究

发布时间：2019-06-26 13:28

【摘要】：紫杉醇(Paclitaxel, PTX)是对多种类型肿瘤细胞都具有重要药理活性的抗肿瘤药物,广泛用于卵巢癌、乳腺癌、非小细胞肺癌、结肠癌、头颈癌、淋巴瘤以及由艾滋病引起的卡波西氏肉瘤等的治疗。紫杉醇的水溶性极低,现临床应用的紫杉醇主要为紫杉醇注射液(商品名为泰素,Taxol),它由聚氧乙烯蓖麻油(Cremophor EL)和无水乙醇(1：1,v/v)配制而成。但是处方中的辅助溶剂,聚氧乙烯蓖麻油,静脉注射时易引起严重的过敏反应,从而限制其应用。因此,大量的研究都致力于紫杉醇的口服制剂的研发以减少静脉注射引起的不良反应。但是,由于紫杉醇水溶性差,同时又是CYP3A4和P-gp的底物,致使其口服时生物利用度很低。紫杉醇脂质处方可以改善紫杉醇的口服生物利用度。脂质处方改善紫杉醇口服生物利用度的机制包括增加药物在胃肠道的溶解度,以及提高紫杉醇的肠淋巴转运。但目前尚未见紫杉醇的肠淋巴转运的相关报道。因此,本文将对紫杉醇的肠淋巴转运进行研究。该研究主要分三个部分进行,具体如下：第一部分：建立大鼠血浆和淋巴中紫杉醇检测的高效液相色谱方法,并对方法进行验证。血浆和淋巴以地西泮为内标,经乙腈沉淀,以Waters Symmetry(?) C18 (150 × 4.6 mm,5μm为色谱柱；流动相为乙腈-水溶液(48：52,v/v),流速为1mL1·min-1;柱温为30℃,检测波长为227 nm。血浆和淋巴样品的进样体积分别为50μL和20μL。淋巴中紫杉醇浓度在005～10μg·mL‘范围内线性关系良好(r=0.9996)。最低定量限为0.05μg·mL-1。低(0.05μg·mL中(0.5 μg·mL-1)高(5μg·mL-1)3个浓度的绝对回收率分别为(78.1±4.65)%、(86.1±9.56)%、(85.2±5.19)%；日内和日间RSD均小于15%。血浆中紫杉醇浓度在0.02～5μg·mL-1范围内线性关系良好(r=0.9999)。最低定量限为0.02μg·mL-1。低(0.05μg·ml-1)、中(0.5μg·mL-1)、高(2μg·mL-1)3个浓度的绝对回收率分别为(77.3±5.61)%、(77.0±2.33)%、(83.1±5.79)%；日内日间精密度RSD均小于10%。该方法准确、灵敏、快速、简便,可适用于紫杉醇在大鼠体内的药动学研究。第二部分：建立麻醉大鼠肠系膜淋巴管、右颈静脉、十二指肠三插管模型,十二指肠分别给予20 mg·kg-1紫杉醇溶液、紫杉醇溶液经P-gp抑制剂维拉帕米预处理、紫杉醇溶液剂与单甘油酯/亚油酸混合物合用、紫杉醇溶液剂经P-gp抑制剂维拉帕米预处理与单甘油酯/亚油酸混合物合用,收集淋巴液和血浆样品进行HPLC测定。结果表明,紫杉醇溶液的累积肠淋巴转运率为(0.038±0.003)%,紫杉醇存在肠淋巴转运现象。紫杉醇溶液与单甘油酯/亚油酸混合物一起服用可使肠淋巴转运程度增加,但是两者血浆中的AUC、Cmax并无显著差异。紫杉醇溶液经维拉帕米预处理,其累积肠淋巴转运率为(0.136±0.022)%,同时绝对生物利用度与对照组相比提高了1.8倍。紫杉醇溶液剂经维拉帕米预处理并和单甘油酯/亚油酸混合物一起服用可使紫杉醇的累积肠淋巴转运率达到(0.245±0.027)%,与对照组相比提高了6.5倍,绝对生物利用度提高了1.8倍。因此,脂质可增加紫杉醇的肠淋巴转运程度,P-gp抑制剂可以同时提高紫杉醇的肠淋巴转运程度以及生物利用度。第三部分：制备紫杉醇自微乳化药物传递系统,对其在大鼠体内的肠淋巴转运程度进行考察,并与紫杉醇溶液剂进行对比。采用伪三元相图,筛选并优化紫杉醇自微乳化药物传递系统的处方。建立麻醉大鼠肠系膜淋巴管、右颈静脉、十二指肠三插管模型,十二指肠给予20 mg·kg-1紫杉醇自微乳制剂,收集淋巴和血浆样品用以测定。紫杉醇自微乳制剂的累积肠淋巴转运率为(0.151±0.022)%,较紫杉醇溶液组有显著提高。另外紫杉醇自微乳制剂的Cmax和AUC均高于溶液剂,生物利用度提高。因此紫杉醇自微乳制剂可用于提高紫杉醇的肠淋巴转运程度及生物利用度。本文的研究结果,有助于阐明口服紫杉醇的肠道吸收转运规律、紫杉醇脂质给药系统口服肠淋巴转运的特点及新型制剂的研发提供实验依据。
[Abstract]:Paclitaxel (PTX) is an anti-tumor drug with important pharmacological activity for various types of tumor cells, and is widely used for the treatment of ovarian cancer, breast cancer, non-small cell lung cancer, colon cancer, head and neck cancer, lymphoma, and Kaposi's sarcoma caused by AIDS. The water-solubility of paclitaxel is very low, and the clinical application of the paclitaxel is mainly paclitaxel injection (the trade name of Taxol, Taxol), which is prepared by the preparation of polyoxyethylene and sesame oil (Cremophor EL) and absolute ethyl alcohol (1:1, v/ v). But the auxiliary solvent, polyoxyethylene and sesame oil in the prescription are easy to cause severe allergic reaction in the intravenous injection, thereby limiting the application. Therefore, a large number of studies are directed to the development of oral formulations of paclitaxel to reduce the adverse reactions caused by intravenous injection. However, due to the poor water solubility of paclitaxel, it is also a substrate of CYP3A4 and P-gp, resulting in low bioavailability when administered orally. The paclitaxel lipid formulation can improve the oral bioavailability of paclitaxel. The mechanism of the lipid formulation to improve the oral bioavailability of paclitaxel includes increasing the solubility of the drug in the gastrointestinal tract, and increasing the intestinal lymphatic transport of the paclitaxel. However, the relevant reports of the intestinal lymphatic transport of paclitaxel have not been reported. Therefore, this paper will study the intestinal lymphatic transport of paclitaxel. The study was carried out in three parts, including the first part: to establish a high performance liquid chromatography method for the detection of paclitaxel in plasma and lymph of rats, and to verify the method. Plasma and lymph were internal standard, precipitated with acetonitrile, with Waters Symmetry (? ) The mobile phase is acetonitrile-water solution (48:52, v/ v), the flow rate is 1 mL1 路 min-1, the column temperature is 30 鈩，

本文编号：2506217