包覆抗癌藥物的MPEG-PCL奈米粒子負載麥胚凝集素與葉酸對人類惡性膠質母細胞瘤的標靶傳遞

本研究合成雙標靶的聚乙二醇聚己內脂奈米粒子(methoxy poly(ethylene glycol)-poly(ε-caprolactone), MPEG-PCL)奈米粒子作為藥物載體，以麥胚凝集素(wheat germ agglutinin)和葉酸(folic acid)修飾於粒子表面，使抗癌藥物能夠穿透血腦屏障並標靶人類惡性膠質母細胞瘤(malignant glioblastoma)。由Pluronic F127 作為界面活性劑以微乳化溶劑揮發法製備奈米粒子，再利用人腦微血管內皮細胞、人類星形細胞及人類周細胞建立體外血腦屏障模型(in vitro blood–brain barrier model)探討不同組態的粒子包覆抗癌藥物對體外血腦屏障的穿透率和雙標靶效應對人類惡性膠質母細胞U87MG cells的抑制，最後藉由免疫螢光染色觀察奈米粒子被人腦微血管內皮細胞和U87MG cells攝入的結果。由實驗結果顯示，奈米粒子的粒徑會隨著界面活性劑濃度由0.25%增加至1%而減少，當MPEG114-PCL40 (PEPC1)製備的奈米粒子在F127濃度為1%時為合成的最佳粒徑，約為150 nm。此外，較短PCL的共聚物可形成較小的粒子、較低的藥物包覆率，但藥物釋放的效果較好。在藥物釋放的結果中表示，本研究製備的粒子具有依賴pH值的釋放行為；由體外血腦屏障模型所測得的transepithelial electrical resistance (TEER)值為310 ± 11.7 Ω × cm2，而各組態的粒子穿透率為FWNPs > WNPs > FNPs > NPs，雙標靶的奈米粒子FWNPs的穿透率最高，約是未改質粒子的1.5~2.1倍，且由雙標靶效應的結果證明FWNP穿透血腦屏障後能夠有效抑制U87MG cells。最後經由免疫螢光染色結果證實雙標靶的奈米粒子能夠增加HBMECs和U87MG cells對奈米粒子的攝入量；因此，以麥胚凝集素和葉酸作為雙標靶的奈米粒子，未來可期待有效應用於腦腫瘤的治療。

關鍵字

血腦屏障；腦癌；聚乙二醇聚己內脂奈米粒子；雙標靶；麥胚凝集素；葉酸

並列摘要

A dual-targeting drug carrier methoxy poly(ethylene glycol)-poly(ε-caprolactone) nanoparticles (MPEG-PCL NPs) and surface modified with wheat germ agglutinin (WGA) and folic acid (FA) for enhanced anticancer drug delivery across the blood–brain barrier (BBB) and targeting to U87MG cells. The MPEG-PCL NPs was prepared using emulsion-solvent evaporation techniques in presence of Pluronic F127. Furthermore, the cell co-culture model was established by consisting of human brain-microvascular endothelial cells, human astrocytes and human brain vascular pericytes to investigate various formulations of drug-loaded NPs permeability of the in vitro BBB model and dual-targeting effect for U87MG cells, then observing the cellular uptake of human brain-microvascular endothelial cells and U87MG cells by immunofluorescence. The results indicated that increase in concentration of F127 from 0.25% to 1% made significant decrease in the average diameter and 1% w/v of F127 was considered as optimum for the preparation of nanoparticles, the optimum average diameter was about 150 nm. In addition, copolymers with shorter PCL blocks formed smaller particles size and encapsulated less drug molecules, but released faster. The results of the release showed that the NPs in this study were pH-dependent. Transepithelial electrical resistance (TEER) value of the in vitro BBB model was 310 ± 11.7 Ω × cm2 and the order of transport across the BBB was FWNPs > WNPs > FNPs > NPs, the dual-targeting nanoparticles FWNPs was 1.5- to 2.1-fold higher than non-modified NPs and the result of dual-targeting effect indicated that FWNPs were significantly against U87MG cells after cross the BBB. Finally, the results of immunofluorescence demonstrated that FWNPs increased intracellular in HBMECs and U87MG cells. Therefore, it may be concluded that FWNPs has the potential to be applied as a targeted delivery for anticancer drug in the treatment of brain cancer.

並列關鍵字

dual-targeting ； blood–brain barrier ； brain cancer ； MPEG-PCL nanoparticles ； wheat germ agglutinin ； folic acid

參考文獻

[1] Bhojani, M. S., Van Dort, M., Rehemtulla, A., Ross, B. D., "Targeted imaging and therapy of brain cancer using theranostic nanoparticles," Mol. Pharm., 7, 1921－1929, 2010.

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[4] Martiny-Baron, G., Marme, D., "Vegf-mediated tumour angiogenesis: A new target for cancer therapy," Curr. Opin. Biotechnol., 6, 675－680, 1995.

[5] Needle, M. N., Molloy, P. T., Geyer, J. R., Herman-Liu, A., Belasco, J. B., Goldwein, J. W., Sutton, L., Phillips, P. C., "Phase ii study of daily oral etoposide in children with recurrent brain tumors and other solid tumors," Med. Pediatr. Oncol., 29, 28－32, 1997.

[6] Stewart, L. A., "Chemotherapy in adult high-grade glioma: A systematic review and meta-analysis of individual patient data from 12 randomised trials," Lancet, 359, 1011－1018, 2002.

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包覆抗癌藥物的MPEG-PCL奈米粒子負載麥胚凝集素與葉酸對人類惡性膠質母細胞瘤的標靶傳遞

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