共载双硫仑与阿霉素纳米胶束的制备和体外评价

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摘要目的：制备共载双硫仑（DSF）与阿霉素（DOX）纳米胶束，评价其体外理化性质以及逆转乳腺癌细胞对DOX耐药的效果。方法：以聚乙二醇单甲醚-聚乳酸共聚羟基乙酸-聚谷氨酸（mPEG-PLGA-PGA）为材料，以透析法制备双载药纳米胶束（DSFDOX-NPs），以动态光散射激光粒度测定仪（DLS）、透射电镜（TEM）对其理化性质进行表征，HPLC测量载药量和包封率，动态透析法测定药物体外释放行为，MTT以及激光共聚焦显微镜评价其逆转人乳腺癌MCF-7/ADR细胞对DOX耐药的效果。结果：DLS测定DSFDOX-NPs平均粒径为（107.5± 0.2）nm，Zeta电位为（-13.7±0.1）mV；TEM显示为球形颗粒，粒径大小为95 nm；HPLC测定DSF载药量为1.91%，包封率为80.30%，DOX载药量为2.28%，包封率为96.2%；体外释放表明DSFDOX-NPs对2种药物均有明显的缓释作用，呈现DSF先快释放、DOX后慢释放的“级联式”释放模式；体外细胞实验表明DSFDOX-NPs能显著地增加DOX在耐药细胞中的浓度，相比DOX溶液剂具有更高的细胞毒性。结论：DSFDOX-NPs有效地装载了2种药物，其粒径小于100 nm，分布均匀，体外缓慢释放药物，并且逆转了DOX耐药肿瘤细胞的耐药效果。

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关键词 ：双硫仑, 盐酸阿霉素, 嵌段聚合物, 胶束, 聚谷氨酸, 聚乳酸共聚羟基乙酸, 乳腺肿瘤, 多药耐药

Abstract：Objective: To prepare the nano-scal polymer micelles simultaneously encapsulating disulfiram (DSF) and doxorubicin (DOX) and to evaluate its physical and chemical properties in vitro and the ability to reverse the multi-drugs resistance of breast cancer cells against doxorubicin. Methods: Using monomethoxy (polyethylene glycol)-b-P (D, L-lactic-co-glycolic acid)-b-P (L-glutamic acid) (mPEG-PLGA-PGA) as materials, the dual-drugs loaded polymer micelles (DSFDOX-NPs) was prepared by dialysis method and its physical and chemical properties were studied by dynamic light scattering (DLS), transmission electron microscope (TEM). Drug loading content (DLC) and drug loading efficiency (DLE) were measured by HPLC. Furthermore, in vitro drug release profiles from DSFDOX-NPs were explored by dynamic dialysis method. Finally, the cellular toxicity against MCF-7/ADR and the cellular uptake of DSFDOX-NPs were evaluated. Results: DSFDOX-NPs had a mean diameter of (107.5±0.2) nm, and Zeta potential of (-13.7±0.1) mV, determined by DLS. TEM image showed that DSFDOX-NPs had spherical shape, and dry mean diameter of 95 nm. DLC and DLE of disulfiram was 1.91% and 80.30% respectively, while DLC and DLE of doxorubicin was 2.28% and 96.2% respectively. The sustained-release of both of DSF and DOX from DSFDOX-NPs was observed, exhibiting a cascade release profiles with the faster release of DSF followed by a slower release of DOX. DSFDOX-NPs can enhance the cellular uptake of DOX by drug-resistant MCF-7/ADR cells through inhibition of P-gP, displaying the significantly higher cytotoxicity against MCF-7/ADR than DOX solution. Conclusion: DSF and DOX is efficiently encapsulated in DSFDOX-NPs with a particle size of less than 100 nm. The sustained-release of DSF and DOX from DSFDOX-NPs is also observed. More importantly, DSFDOX-NPs displayed multidrugresistance reversal effect on MCF-7/ADR cell.

Key words： disulfiram doxorubicin block copolymer micelles polyglutamicacid polylactic-co-glycolic acid breast neoplasms multi-drugs resistanc

收稿日期: 2017-08-05

基金资助:浙江省大学生科技创新活动计划暨新苗人才计划项目（2017R413059）；国家自然科学基金青年基金资助项目（81603036）。

通讯作者: 徐荷林，硕士生导师，Email：xhlpharm1214@126.com

作者简介: 徐霞芳（1995-），女，浙江嵊州人，本科生。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2018/V48/I2/79

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