Cheng Xinwei, Liu Qibing, Li Hong, Kang Chen, Liu Yang, Guo Tianqi, Shang Ke, Yan Chengyun, Cheng Guang, Lee Robert J
Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio, 43210, USA.
Department of Pharmacology, Hainan Medical University, Xueyuan Road, Haikou, 571199, Hainan, China.
Pharm Res. 2017 Feb;34(2):310-320. doi: 10.1007/s11095-016-2063-5. Epub 2016 Nov 28.
Bcl-2 is an anti-apoptotic gene that is frequently overexpressed in human cancers. G3139 is an antisense oligonucleotide against bcl-2 that has shown limited efficacy in clinical trials. Here, we report the synthesis of a new antisense oligonucleotide containing additional chemical modifications and its delivery using nanoparticles.
An oligonucleotide G3139-GAP was synthesized, which has 2'-O-methyl nucleotides at the 5' and 3' ends based on a "gapmer" design. Furthermore, G3139-GAP was incorporated into lipid nanoparticles (LNPs) composed of DOTAP/egg PC/cholesterol/Tween 80. The LNP-loaded G3139-GAP was evaluated in A549 lung cancer cells both in vitro and in a murine xenograft model for biological activity and therapeutic efficacy.
The LNPs showed excellent colloidal and serum stability, and high encapsulation efficiency for G3139-GAP. They have a mean particle diameter and zeta potential of 134 nm and 9.59 mV, respectively. G3139-GAP-LNPs efficiently downregulated bcl-2 expression in A549 cells, as shown by 40% and 83% reduction in mRNA and protein levels, respectively. Furthermore, G3139-GAP-LNPs were shown to inhibit tumor growth, prolong survival, and downregulate tumor bcl-2 expression in an A549 murine xenograft tumor model. These data indicate that G3139-GAP-LNPs have excellent anti-tumor efficacy and warrant further evaluation.
Bcl-2是一种抗凋亡基因，在人类癌症中经常过度表达。G3139是一种针对bcl-2的反义寡核苷酸，在临床试验中显示出有限的疗效。在此，我们报告一种含有额外化学修饰的新型反义寡核苷酸的合成及其使用纳米颗粒的递送。
合成了一种寡核苷酸G3139-GAP，基于“gapmer”设计，其5'和3'端具有2'-O-甲基核苷酸。此外，G3139-GAP被掺入由DOTAP/蛋黄卵磷脂/胆固醇/吐温80组成的脂质纳米颗粒（LNP）中。在A549肺癌细胞中对负载LNP的G3139-GAP进行体外和小鼠异种移植模型评估，以检测其生物活性和治疗效果。
LNP显示出优异的胶体稳定性和血清稳定性，以及对G3139-GAP的高包封效率。它们的平均粒径和zeta电位分别为134 nm和9.59 mV。如mRNA和蛋白质水平分别降低40%和83%所示，G3139-GAP-LNP在A549细胞中有效下调bcl-2表达。此外，在A549小鼠异种移植肿瘤模型中，G3139-GAP-LNP显示出抑制肿瘤生长、延长生存期和下调肿瘤bcl-2表达的作用。这些数据表明G3139-GAP-LNP具有优异的抗肿瘤疗效，值得进一步评估。
