Department of Pharmacology, School of Basic Medicine and Life Science , Hainan Medical University , Haikou , China.
State Key Laboratory of Long-Acting and Targeted Drug Delivery, Nanjing , China.
Mol Pharm. 2018 Oct 1;15(10):4722-4732. doi: 10.1021/acs.molpharmaceut.8b00696. Epub 2018 Sep 7.
Expression of Bcl-2 and Akt-1 has been associated with human cancer. G3139 and RX-0201, targeting Bcl-2 and Akt-1, respectively, are antisense oligonucleotides (ASOs) that have shown limited efficacy in clinical trials. Herein, we report a combination of newly designed ASOs based on these agents and was delivered by tumor cell-targeting lipid nanoparticles (LNPs). A "Gapmer" design strategy was applied to these ASOs with the addition of 2'-O-methyl modifications on the nucleotides at 5' and 3' ends. A dual-channel syringe pump-based system was developed for the synthesis of the LNPs. ASO-LNPs composed of DODMA, egg PC, cholesterol, T7-PEG-DSPE, and PEG-DMG at a molar ratio of 35:39.5:20:0.5:5 and carrying either individual ASOs or co-loaded ASO combinations (Co-ASOs) were synthesized and evaluated in both KB and A549 cancer cells and in an A549 murine xenograft model to determine their antitumor effects and biological activities. The ASO-LNPs exhibited excellent colloidal stability and high ASO encapsulation efficiency with relatively small mean particle sizes and moderately positive zeta potentials. Transferrin receptor-targeting T7-conjugated LNPs showed enhanced cellular uptake compared to nontargeted LNPs. In addition, both T7-conjugated Co-ASOs-LNPs and non-T7-conjugated Co-ASOs-LNPs at a molar ratio of (G3139-GAP to RX-0201-GAP at 1:2) showed efficient downregulation of both Bcl-2 and Akt-1 in both A549 and KB cells. Furthermore, T7-conjugated Co-ASOs-LNPs (Co-ASOs-LNPs) produced superior antitumor activity, prolonged the overall survival time, and demonstrated tumor targeting activity in an A549 xenograft model.
Bcl-2 和 Akt-1 的表达与人类癌症有关。分别针对 Bcl-2 和 Akt-1 的 G3139 和 RX-0201 是反义寡核苷酸 (ASO)，它们在临床试验中显示出有限的疗效。在此，我们报告了一种基于这些药物的新设计 ASO 的组合，并通过肿瘤细胞靶向脂质纳米粒 (LNP) 进行递送。“Gapmer”设计策略被应用于这些 ASO，在核苷酸的 5' 和 3' 端添加了 2'-O-甲基修饰。开发了一种双通道注射器泵系统用于 LNP 的合成。由 DODMA、蛋黄 PC、胆固醇、T7-PEG-DSPE 和 PEG-DMG 以摩尔比 35:39.5:20:0.5:5 组成的 ASO-LNP，并携带单独的 ASO 或共载 ASO 组合 (Co-ASO) 进行合成和评估，在 KB 和 A549 癌细胞中和 A549 小鼠异种移植模型中，以确定它们的抗肿瘤作用和生物学活性。ASO-LNP 表现出优异的胶体稳定性和高 ASO 包封效率，具有相对较小的平均粒径和适度正的 ζ 电位。与非靶向 LNP 相比，转铁蛋白受体靶向 T7 缀合的 LNP 显示出增强的细胞摄取。此外，T7 缀合的 Co-ASO-LNP 和非 T7 缀合的 Co-ASO-LNP 的摩尔比（G3139-GAP 与 RX-0201-GAP 为 1:2）均能有效下调 A549 和 KB 细胞中的 Bcl-2 和 Akt-1。此外，T7 缀合的 Co-ASO-LNP (Co-ASO-LNP) 在 A549 异种移植模型中产生了优异的抗肿瘤活性，延长了总生存时间，并表现出肿瘤靶向活性。
