Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. Albert Hall, B49, St. John's University, 8000 Utopia Parkway, Queens, New York, 11439, USA.
Pharm Res. 2022 Nov;39(11):2745-2759. doi: 10.1007/s11095-022-03184-3. Epub 2022 Feb 10.
The present work was aimed at developing an optimized and modified nanostructured lipid carrier of BRD4 protein degrading Proteolysis Targeting Chimera (PROTAC) against non-small cell lung carcinoma. PROTACs are an emerging class of anticancer molecules with nanomolar activity but associated with significant solubility challenges. Lipid-based colloidal systems like nanostructured lipid carriers are widely explored for such highly lipophilic molecules. ARV-825, a cereblon-based PROTAC was investigated for its anticancer efficacy in vitro in 2D and 3D lung cancer models. ARV-825 loaded PEGylated nanostructured lipid carriers (AP-NLC) was prepared using melt emulsification technique. ARV-825 was stabilized using Precirol® ATO5 and Captex® 300 EP/NF as the solid and liquid lipid, respectively. However, hydrophobic ion-pairing with medium chain fatty acid was required to improve drug loading and stability. A hydrodynamic diameter and polydispersity index of 56.33 ± 0.42 nm and 0.16 respectively with zeta potential of -21 ± 1.24 mV was observed. In vitro migration and colony formation assay confirmed the anticancer activity of ARV-825 alone and AP-NLC. Nearly 38% and 50% apoptotic cell population were observed after ARV-825 and AP-NLC treatment. Immunoblotting assay showed complete suppression of BRD4 and c-Myc protein expression for AP-NLC. Most importantly, significant reduction in the growth of multicellular 3D spheroid of A549 cells confirmed the effectiveness of BRD4 PROTAC and its lipid nanoparticle in non-small cell lung cancer (NSCLC). AP-NLC. Higher amount of red fluorescence throughout the spheroid surface further confirmed superior efficacy of AP-NLC in tumor penetration and cell killing.
本工作旨在开发一种针对非小细胞肺癌的 BRD4 蛋白降解 PROTAC 的优化和改良的纳米结构脂质载体。PROTAC 是一类新兴的抗癌分子，具有纳摩尔级的活性，但与显著的溶解度挑战相关。基于脂质的胶体系统，如纳米结构脂质载体，广泛用于此类高度亲脂性分子。ARV-825 是一种基于 cereblon 的 PROTAC，已在 2D 和 3D 肺癌模型中进行了体外抗癌功效的研究。使用熔融乳化技术制备 ARV-825 负载的 PEG 化纳米结构脂质载体（AP-NLC）。使用 Precirol® ATO5 和 Captex® 300 EP/NF 分别作为固体和液体脂质来稳定 ARV-825。然而，需要中等链脂肪酸的疏水离子配对来提高药物载药量和稳定性。观察到 56.33±0.42nm 的水动力学直径和 0.16 的多分散指数，zeta 电位为-21±1.24mV。体外迁移和集落形成试验证实了 ARV-825 单独和 AP-NLC 的抗癌活性。在用 ARV-825 和 AP-NLC 处理后，观察到近 38%和 50%的凋亡细胞群体。免疫印迹分析显示，AP-NLC 完全抑制了 BRD4 和 c-Myc 蛋白的表达。最重要的是，A549 细胞的多细胞 3D 球体的生长显著减少，证实了 BRD4 PROTAC 及其脂质纳米颗粒在非小细胞肺癌（NSCLC）中的有效性。AP-NLC。球体表面的红色荧光量增加进一步证实了 AP-NLC 在肿瘤渗透和细胞杀伤方面的优越疗效。
