Department of Pharmaceutics, Faculty of Pharmacy, Kalabhavan Campus, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat 390001, India.
Shobhaben Pratapbhai Patel School of Pharmacy & Technology Management, SVKM's NMIMS University, Mumbai, Maharashtra, 400056, India.
Drug Deliv Transl Res. 2021 Oct;11(5):2052-2071. doi: 10.1007/s13346-020-00867-5. Epub 2020 Nov 11.
The resistance of cancer cells to chemotherapy has presented a formidable challenge. The current research aims at evaluating whether silencing of the cisplatin efflux promoter gene ABCC3 using siRNA co-loaded with the drug in a nanocarrier improves its efficacy in non-small cell lung cancer (NSCLC). Hybrid nanocarriers (HNCs) comprising lipids and poly(lactic acid-polyethylene glycol) di-block copolymer (PEG-PLA) were prepared for achieving the simultaneous delivery of cisplatin caprylate and ABCC3-siRNA to the cancer cells. PEGylation of the formulated HNCs was carried out using post-insertion technique for imparting long circulation characteristics to the carrier. The optimized formulation exhibited an entrapment efficiency of 71.9 ± 2.2% and 95.83 ± 0.39% for cisplatin caprylate and siRNA respectively. Further, the HNC was found to have hydrodynamic diameter of 153.2 ± 1.76 nm and + 25.39 ± 0.49 mV zeta potential. Morphological evaluation using cryo transmission electron microscopy confirmed the presence of lipid bilayer surrounding the polymeric core in HNCs. The in vitro cellular uptake studies showed improved uptake, while cell viability studies of the co-loaded formulation in A549 cell-line indicated significantly improved cytotoxic potential when compared with drug solution and drug-loaded HNCs; cell cycle analysis indicated increased percentage of cell arrest in G2-M phase compared with drug-loaded HNCs. Further, the gene knock-down study showed that silencing of ABCC3 mRNA might be improved in vitro efficacy of the formulation. The optimized cisplatin and ABCC3 siRNA co-loaded formulation presented significantly increased half-life and tumour regression in A549 xenograft model in BALB/c nude mice. In conclusion, siRNA co-loaded formulation presented reduced drug resistance and increased efficacy, which might be promising for the current cisplatin-based treatments in NSCLC.
癌细胞对化疗的耐药性是一个巨大的挑战。本研究旨在评估使用载药纳米载体共载顺铂前药和 ABCC3-siRNA 沉默 ABCC3 基因能否提高其在非小细胞肺癌(NSCLC)中的疗效。采用脂质和聚乳酸-聚乙二醇二嵌段共聚物(PEG-PLA)制备混合纳米载体(HNC),实现顺铂辛酸酯和 ABCC3-siRNA 同时递送至癌细胞。采用后插入技术对 HNC 进行 PEG 化,赋予载体长循环特性。优化后的配方对顺铂辛酸酯和 siRNA 的包封效率分别为 71.9±2.2%和 95.83±0.39%。此外,HNC 的水动力学直径为 153.2±1.76nm,+25.39±0.49mV 表面电势。使用冷冻透射电子显微镜进行形态评估证实了 HNC 中存在脂质双层围绕聚合物核。体外细胞摄取研究表明,与药物溶液和载药 HNC 相比,共载体制剂的摄取得到了改善;在 A549 细胞系中进行的共载体制剂细胞活力研究表明,与载药 HNC 相比,其细胞毒性潜力显著提高;细胞周期分析表明,与载药 HNC 相比,G2-M 期的细胞阻滞百分比增加。此外,基因敲低研究表明,ABCC3 mRNA 的沉默可能会提高该制剂的体外疗效。优化后的顺铂和 ABCC3 siRNA 共载体制剂在 BALB/c 裸鼠的 A549 异种移植模型中显著延长了半衰期并促进肿瘤消退。总之,共载体制剂降低了耐药性并提高了疗效,有望为当前基于顺铂的 NSCLC 治疗提供新的思路。