This study aimed to design an effective targeted combination of doxorubicin (Dox)-Curcumin (Cur) delivery system to eradicate the MDA-MB231 cell line. A novel biodegradable poly ε-Caprolactone-co-maleic anhydride-graft-citric acid copolymer micelle (PCL-co-P(MA-g-CA)) was synthesized through thiolen radical copolymerization and ring-opening polymerization. The unique micelle structure allowed simultaneous loading of hydrophilic Dox and hydrophobic Cur with a loading efficiency of above 98 % for each drug. The physicochemical characterization of copolymeric micelle was analyzed by ¹HNMR, ¹³CNMR, FTIR, DSC, CMC, DLS and SEM. The in vitro cytotoxicity was assessed by MTT assay, cell cycle analysis, annexin V-FITC apoptosis, qRT-PCR and western blot. The final obtained micelles with critical micelle concentration (CMC) of 0.5 μg/mL, and particle size and surface charge was 60 nm and -14.1 mV, respectively. Beside the fast uptake of designed micelle, Dox@Cur loaded micelle showed a synergistic effect with the combination index (CI) value of below 1. Our results revealed that this novel engineered combinatorial micelle induced apoptosis (96 %) which was proved by annexin V and cell cycle. qRT-PCR and western blot assays demonstrated involvement of intrinsic apoptosis pathways in the genetic and protein levels. Finally, the penetration of Dox@Cur loaded micelle was evaluated by 3D in vitro tumor formation. Our findings showed the penetration behavior of micelles is in a concentration-dependent manner. In conclusion, combinational therapy by using Dox and Cur nano-formulation has boosted the cytotoxicity in MDA-MB231 cells by promoting the apoptotic response.

这项研究旨在设计一种有效的靶向性阿霉素（Dox）-姜黄素（Cur）递送系统组合，以根除MDA-MB231细胞系。通过巯基自由基共聚和开环聚合反应，合成了一种新型的可生物降解的聚ε-己内酯-马来酸酐-接枝-柠檬酸共聚物胶束（PCL-co-P（MA-g-CA））。独特的胶束结构允许同时装载亲水性Dox和疏水性Cur，每种药物的装载效率均高于98％。共聚胶束的理化特性通过¹ HNMR，¹³CNMR，FTIR，DSC，CMC，DLS和SEM。通过MTT测定，细胞周期分析，膜联蛋白V-FITC凋亡，qRT-PCR和western blot评估体外细胞毒性。最终获得的胶束的临界胶束浓度（CMC）为0.5μg/ mL，粒径和表面电荷分别为60 nm和-14.1 mV。除快速吸收设计的胶束外，Dox @ Cur负载的胶束还具有协同作用，其组合指数（CI）值低于1。我们的结果表明，这种新型工程改造的组合胶束可诱导细胞凋亡（96％），这由Annexin V证明。和细胞周期。qRT-PCR和western blot分析表明内在的凋亡途径参与了基因和蛋白质水平。最后，通过3D体外肿瘤形成评估了Dox @ Cur胶束的渗透。我们的发现表明，胶束的渗透行为是浓度依赖性的。总之，使用Dox和Cur纳米制剂的联合疗法通过促进细胞凋亡反应，增强了MDA-MB231细胞的细胞毒性。