Purpose

To develop a new nanoparticle formulation for a proteasome inhibitor Carfilzomib (CFZ) to improve its stability and efficacy for future in vivo applications.

Methods

CFZ-loaded ternary polypeptide nanoparticles (CFZ/tPNPs) were prepared by using heptakis(6-amino-6-deoxy)-β-cyclodextrin(hepta-hydrochloride) (HaβCD) and azido-poly(ethylene glycol)-block-poly(L-glutamic acid sodium salt) (N₃-PEG-PLE). The process involved ternary (hydrophobic/ionic/supramolecular) interactions in three steps: 1) CFZ was entrapped in the cavity of HaβCD by hydrophobic interaction, 2) the drug-cyclodextrin inclusion complexes were mixed with N₃-PEG-PLE to form polyion complex nanoparticles, and 3) the nanoparticles were modified with fluorescent dyes (AFDye 647) for imaging and/or epithelial cell adhesion molecule (EpCAM) antibodies for cancer cell targeting. CFZ/tPNPs were characterized for particle size, surface charge, drug release, stability, intracellular uptake, proteasome inhibition, and in vitro cytotoxicity.

Results

tPNPs maintained an average particle size of 50 nm after CFZ entrapment, EpCAM conjugation, and freeze drying. tPNPs achieved high aqueous solubility of CFZ (>1 mg/mL), sustained drug release (t_1/2 = 6.46 h), and EpCAM-mediated cell targeting, which resulted in increased intracellular drug accumulation, prolonged proteasome inhibition, and enhanced cytotoxicity of CFZ in drug-resistant DLD-1 colorectal cancer cells.

Conclusions

tPNPs improved stability and efficacy of CFZ in vitro, and these results potentiate effective cancer treatment using CFZ/tPNPs in future vivo studies.

中文翻译：

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具有改进的卡非佐米封装，持续释放和增强的体外功效的三元多肽纳米颗粒

目的

开发用于蛋白酶体抑制剂卡非佐米（CFZ）的新纳米颗粒制剂，以提高其稳定性和功效，以用于未来的体内应用。

方法

负载CFZ的三元多肽纳米粒子（CFZ / tPNPs）是通过使用七（6-氨基-6-脱氧）-β-环糊精（庚二盐酸盐）（HaβCD）和叠氮基-聚（乙二醇）-嵌段-聚（ L-谷氨酸钠盐）（N ₃ -PEG-PLE）。该过程涉及三步（疏水/离子/超分子）相互作用：1）CFZ通过疏水相互作用被包埋在HaβCD的空腔中； 2）药物-环糊精包合物与N ₃混合-PEG-PLE形成聚离子复合纳米颗粒，并且3）用荧光染料（AFDye 647）修饰纳米颗粒以成像和/或上皮细胞粘附分子（EpCAM）抗体用于癌细胞靶向。CFZ / tPNPs具有粒径，表面电荷，药物释放，稳定性，细胞内摄取，蛋白酶体抑制和体外细胞毒性的特征。

结果

在CFZ截留，EpCAM偶联和冷冻干燥后，tPNP的平均粒径保持50 nm。tPNP实现了CFZ的高水溶性（> 1 mg / mL），持续药物释放（t _1/2  = 6.46 h）和EpCAM介导的细胞靶向作用，从而导致细胞内药物积累增加，蛋白酶体抑制时间延长和细胞毒性增强CFZ在耐药性DLD-1大肠癌细胞中的作用

结论

tPNPs改善了CFZ的体外稳定性和功效，这些结果增强了CFZ / tPNPs在未来体内研究中的有效癌症治疗效果。