發(fā)布時(shí)間：2019-06-26 13:28

【摘要】：紫杉醇(Paclitaxel, PTX)是對(duì)多種類型腫瘤細(xì)胞都具有重要藥理活性的抗腫瘤藥物,廣泛用于卵巢癌、乳腺癌、非小細(xì)胞肺癌、結(jié)腸癌、頭頸癌、淋巴瘤以及由艾滋病引起的卡波西氏肉瘤等的治療。紫杉醇的水溶性極低,現(xiàn)臨床應(yīng)用的紫杉醇主要為紫杉醇注射液(商品名為泰素,Taxol),它由聚氧乙烯蓖麻油(Cremophor EL)和無水乙醇(1：1,v/v)配制而成。但是處方中的輔助溶劑,聚氧乙烯蓖麻油,靜脈注射時(shí)易引起嚴(yán)重的過敏反應(yīng),從而限制其應(yīng)用。因此,大量的研究都致力于紫杉醇的口服制劑的研發(fā)以減少靜脈注射引起的不良反應(yīng)。但是,由于紫杉醇水溶性差,同時(shí)又是CYP3A4和P-gp的底物,致使其口服時(shí)生物利用度很低。紫杉醇脂質(zhì)處方可以改善紫杉醇的口服生物利用度。脂質(zhì)處方改善紫杉醇口服生物利用度的機(jī)制包括增加藥物在胃腸道的溶解度,以及提高紫杉醇的腸淋巴轉(zhuǎn)運(yùn)。但目前尚未見紫杉醇的腸淋巴轉(zhuǎn)運(yùn)的相關(guān)報(bào)道。因此,本文將對(duì)紫杉醇的腸淋巴轉(zhuǎn)運(yùn)進(jìn)行研究。該研究主要分三個(gè)部分進(jìn)行,具體如下：第一部分：建立大鼠血漿和淋巴中紫杉醇檢測(cè)的高效液相色譜方法,并對(duì)方法進(jìn)行驗(yàn)證。血漿和淋巴以地西泮為內(nèi)標(biāo),經(jīng)乙腈沉淀,以Waters Symmetry(?) C18 (150 × 4.6 mm,5μm為色譜柱；流動(dòng)相為乙腈-水溶液(48：52,v/v),流速為1mL1·min-1;柱溫為30℃,檢測(cè)波長(zhǎng)為227 nm。血漿和淋巴樣品的進(jìn)樣體積分別為50μL和20μL。淋巴中紫杉醇濃度在005～10μg·mL‘范圍內(nèi)線性關(guān)系良好(r=0.9996)。最低定量限為0.05μg·mL-1。低(0.05μg·mL中(0.5 μg·mL-1)高(5μg·mL-1)3個(gè)濃度的絕對(duì)回收率分別為(78.1±4.65)%、(86.1±9.56)%、(85.2±5.19)%；日內(nèi)和日間RSD均小于15%。血漿中紫杉醇濃度在0.02～5μg·mL-1范圍內(nèi)線性關(guān)系良好(r=0.9999)。最低定量限為0.02μg·mL-1。低(0.05μg·ml-1)、中(0.5μg·mL-1)、高(2μg·mL-1)3個(gè)濃度的絕對(duì)回收率分別為(77.3±5.61)%、(77.0±2.33)%、(83.1±5.79)%；日內(nèi)日間精密度RSD均小于10%。該方法準(zhǔn)確、靈敏、快速、簡(jiǎn)便,可適用于紫杉醇在大鼠體內(nèi)的藥動(dòng)學(xué)研究。第二部分：建立麻醉大鼠腸系膜淋巴管、右頸靜脈、十二指腸三插管模型,十二指腸分別給予20 mg·kg-1紫杉醇溶液、紫杉醇溶液經(jīng)P-gp抑制劑維拉帕米預(yù)處理、紫杉醇溶液劑與單甘油酯/亞油酸混合物合用、紫杉醇溶液劑經(jīng)P-gp抑制劑維拉帕米預(yù)處理與單甘油酯/亞油酸混合物合用,收集淋巴液和血漿樣品進(jìn)行HPLC測(cè)定。結(jié)果表明,紫杉醇溶液的累積腸淋巴轉(zhuǎn)運(yùn)率為(0.038±0.003)%,紫杉醇存在腸淋巴轉(zhuǎn)運(yùn)現(xiàn)象。紫杉醇溶液與單甘油酯/亞油酸混合物一起服用可使腸淋巴轉(zhuǎn)運(yùn)程度增加,但是兩者血漿中的AUC、Cmax并無顯著差異。紫杉醇溶液經(jīng)維拉帕米預(yù)處理,其累積腸淋巴轉(zhuǎn)運(yùn)率為(0.136±0.022)%,同時(shí)絕對(duì)生物利用度與對(duì)照組相比提高了1.8倍。紫杉醇溶液劑經(jīng)維拉帕米預(yù)處理并和單甘油酯/亞油酸混合物一起服用可使紫杉醇的累積腸淋巴轉(zhuǎn)運(yùn)率達(dá)到(0.245±0.027)%,與對(duì)照組相比提高了6.5倍,絕對(duì)生物利用度提高了1.8倍。因此,脂質(zhì)可增加紫杉醇的腸淋巴轉(zhuǎn)運(yùn)程度,P-gp抑制劑可以同時(shí)提高紫杉醇的腸淋巴轉(zhuǎn)運(yùn)程度以及生物利用度。第三部分：制備紫杉醇自微乳化藥物傳遞系統(tǒng),對(duì)其在大鼠體內(nèi)的腸淋巴轉(zhuǎn)運(yùn)程度進(jìn)行考察,并與紫杉醇溶液劑進(jìn)行對(duì)比。采用偽三元相圖,篩選并優(yōu)化紫杉醇自微乳化藥物傳遞系統(tǒng)的處方。建立麻醉大鼠腸系膜淋巴管、右頸靜脈、十二指腸三插管模型,十二指腸給予20 mg·kg-1紫杉醇自微乳制劑,收集淋巴和血漿樣品用以測(cè)定。紫杉醇自微乳制劑的累積腸淋巴轉(zhuǎn)運(yùn)率為(0.151±0.022)%,較紫杉醇溶液組有顯著提高。另外紫杉醇自微乳制劑的Cmax和AUC均高于溶液劑,生物利用度提高。因此紫杉醇自微乳制劑可用于提高紫杉醇的腸淋巴轉(zhuǎn)運(yùn)程度及生物利用度。本文的研究結(jié)果,有助于闡明口服紫杉醇的腸道吸收轉(zhuǎn)運(yùn)規(guī)律、紫杉醇脂質(zhì)給藥系統(tǒng)口服腸淋巴轉(zhuǎn)運(yùn)的特點(diǎn)及新型制劑的研發(fā)提供實(shí)驗(yàn)依據(jù)。
[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，

本文編號(hào)：2506217