利用熱敏感性凝膠及混合微胞體為載體傳輸Doxorubicin和Docetaxel於腫瘤之研究

Studies on Drug Delivery of Doxorubicin and Docetaxel to Tumor Using Thermosensitive Hydrogel and Mixed Micelle as Carriers

詹華景(Hua-Jing Jhan)

指導教授 ： 何秀娥

共同指導教授 ： 許明照

臺北醫學大學/藥學院/藥學系(碩博士班)/博士(2015年)

摘要

本研究開發出三種不同的抗癌藥物載體，第一種為含親水性抗癌藥物doxorubicin (Dox) 的可注射熱敏感性凝膠劑型，其最佳處方DH700KMF-15包含1 mg/mL Dox，0.1% Hyaluronic acid (HA700K)，0.1 M MgCl2 以及15% Pluronic F127，製程以簡單的物理性混合法即可製備。第二種為含親脂性抗癌藥物docetaxel (Doc) 的混合微胞體劑型，其最佳處方M-DocLF為Doc，Pluronic L121及Pluronic F127 (1:45:15, w/w) 則以薄膜水合法製備。第三種藥物載體為結合兩種抗癌藥物 (Dox及Doc) 的可注射熱敏感性凝膠，其最佳處方DH700KMF-13.5/M-DocLF組成為1 mg/mL Dox，1 mg/mL Doc，0.1% HA700K，0.1 M MgCl2，13.5% Pluronic F127，45 mg/mL Pluronic L121以及15 mg/mL Pluronic F127。

此三種抗癌藥物載體的物化特性分述如下，流變學試驗顯示DH700KMF-15以及DH700KMF-13.5/M-DocLF在室溫下為可注射的液體而在體溫下便形成不流動的凝膠。M-DocLF混合微胞體的粒徑大小為215.6 ± 11.46 nm，PI為0.286 ± 0.029，表面電荷為 -35.93 ± 4.88 mV，藥物包覆率為98.97%，藥物乘載量為1.62%。腫瘤細胞毒性試驗顯示此三種抗癌藥物載體對於C26結腸癌細胞，HT29人類結腸癌細胞以及MCF7人類乳癌細胞有明顯的抗癌活性。在動物實驗中，以靜脈給予M-DocLF混合微胞體或者將DH700KMF-15或DH700KMF-13.5/M-DocLF凝膠直接注射在動物腫瘤上時，可以有效的抑制Balb/c小鼠上的C26大腸癌細胞生長。

本研究開發出的混合微胞體處方M-DocLF具有被動傳輸特性，可經由靜脈注射給予後藉由通透性增強及停滯效應 (EPR effect) 可聚集於腫瘤附近進而抑制癌細胞。而包含了HA700K的可注射性熱敏感凝膠 (DH700KMF-15以及DH700KMF-13.5/M-DocLF) 則具有專一性標靶局部抑制腫瘤效果，並且能藉由淋巴靶向抑制早期性腫瘤細胞轉移。在動物實驗中證實，最佳的合併Dox以及Doc的可注射性熱敏感凝膠 (DH700KMF-13.5/M-DocLF) 相較於單獨使用Dox或者Doc有顯著的抑制腫瘤效果。藉由藥物動力學及藥物分佈證實本研究所開發劑型釋放出的藥物能夠聚集在腫瘤位置並且減少全身性的副作用產生。

關鍵字

Doxorubicin ； Docetaxel ； 熱敏感性凝膠 ； 混合微胞體 ； 組合療法

並列摘要

In this study, we developed three drug carriers to deliver anticancer drugs. The first formulation is a thermosensitive injectable hydrogel composed of 1 mg/mL Dox, 0.1% Hyaluronic acid (HA700K), 0.1 M MgCl2 and 15% Pluronic F127 designed as DH700KMF-15 was prepared simply by physically mixing. The second formulation is a mixed micelle encapsulated hydrophobic anticancer drug such as docetaxel (Doc). The optimal formulation of M-DocLF contained Doc, Pluronic L121 and Pluronic F127 (1:45:15, w/w) was prepared using a thin film hydration method. The third formulation combination of Dox and Doc were loaded in a thermosensitive injectable hydrogel, DH700KMF-13.5 / M-DocLF, that contained 1 mg/mL Dox, 1 mg/mL Doc, 0.1% HA700K, 0.1 M MgCl2, 13.5% Pluronic F127, 45 mg/mL Pluronic L121 and 15 mg/mL Pluronic F127.

Their physicochemical properties were characterized below : rheology tests showed that DH700KMF-15 and DH700KMF-13.5 / M-DocLF are injectable flowing solutions at room temperature, and form non-flowing gels in body temperature. The particle size, poydispersity index, zeta potential, encapsulation efficiency, and drug loading of M-DocLF mixed micelle was 215.6 ± 11.46 nm, 0.286 ± 0.029, -35.93 ± 4.88 mV, 98.97% and 1.62%, respectively. Cytotoxicity assay revealed that all of them have significantly anti-cancer activities to C26 colon cancers cell, HT29 human colon cancer cell and MCF7 human breast cancer cells. In animal studies, By intravenous administration of M-DocLF or intratumoral administration of DH700KMF-15 or DH700KMF-13.5/ M-DocLF resulted in efficient growth inhibition of C26 colon cancer cell on Balb/c mice model.

In this study, M-DocLF mixed micelle has passive targeting property and it could accumulation in tumor site via enhanced permeability and retention (EPR) effect after intravenous injection. The injectable thermosensitive hydrogel (DH700KMF-15 and DH700KMF-13.5/ M-DocLF) contained HA700K could active targeting, local treatment of cancer disease and lymphotropic targeting to inhibit early metastasis. In animal studies demonstrated the optimally developed combination of Dox and Doc (DH700KMF-13.5/ M-DocLF) showed significantly superior to Dox or Doc when given alone. Pharmacokinetic and biodistribution studies demonstrated drugs released from our formulations could accumulation in tumor site and reduced systemic side effect.

並列關鍵字

Doxorubicin ； Docetaxel ； Thermosensitive hydrogel ； Mixed micelle ； Combination therapy.

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