The Institute for Drug Research, The School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, POB 12065, Jerusalem, 91120, Israel.
Small. 2013 Dec 20;9(24):4221-36. doi: 10.1002/smll.201301417. Epub 2013 Jul 21.
Aberrant signaling of the epidermal growth factor receptor (EGFR) is common to a variety of human cancers and is also found to be over-expressed in most cases of non-small cell lung cancer. For the development of a molecularly targeted therapy, cetuximab-conjugated nanoparticles (immunonanoparticles, INPs) are designed and loaded with the lipophilic paclitaxel palmitate (pcpl) prodrug. Oleyl cysteineamide (OCA) is synthesized whereby its amphiphilic nature enables interfacial anchoring and thiol surface functionalization of PLGA NPs, facilitating bioconjugation to cetuximab by thioether bonds. It is demonstrated that the in vitro targeting efficiency and improved cellular internalization and cytotoxicity of this targeted delivery system in lung cancer cells over-expressing EGFR. A quantitative measure of the high binding affinity of INPs to EGFR is demonstrated using surface plasmon resonance. In vivo tolerability and enhanced efficacy of cetuximab pcpl INPs in a metastatic lung cancer model are reported. Its therapeutic efficacy in A549-luc-C8 lung tumors is shown using non-invasive bioluminescent imaging. Intravenous administration of cetuximab pcpl INPs to mice results in significantly higher inhibition of tumor growth and increased survival rates as compared to the non-targeted drug solution, drug-loaded nanoparticles or blank INPs. Pharmacokinetics and organ biodistribution of the prodrug and parent drug are evaluated by LC-MS/MS in lung tumor bearing mice. No enhanced total accumulation of nanoparticles or INPs is found at the tumor tissue. However, persistent pcpl levels with sustained conversion and release of paclitaxel are observed for the encapsulated prodrug possibly suggesting the formation of a drug reservoir. The overall results indicate the potential of this promising targeted platform for the improved treatment of lung cancer and other EGFR positive tumors.
表皮生长因子受体 (EGFR) 的异常信号传导常见于多种人类癌症，并且在大多数非小细胞肺癌病例中也发现过度表达。为了开发一种分子靶向治疗，设计了与西妥昔单抗缀合的纳米颗粒（免疫纳米颗粒，INPs）并负载疏水性紫杉醇棕榈酸酯（pcpl）前药。合成了油酰半胱氨酸酰胺（OCA），其两亲性质能够使 PLGA NPs 的界面锚定和巯基表面功能化，从而通过硫醚键促进与西妥昔单抗的生物缀合。结果表明，这种靶向递药系统在 EGFR 过表达的肺癌细胞中具有体外靶向效率和改善的细胞内化和细胞毒性。使用表面等离子体共振证明了 INPs 与 EGFR 的高结合亲和力的定量测量。报告了转移性肺癌模型中西妥昔单抗 pcpl INPs 的体内耐受性和增强的疗效。使用非侵入性生物发光成像显示了 A549-luc-C8 肺肿瘤中 INPs 的治疗效果。与非靶向药物溶液、载药纳米颗粒或空白 INPs 相比，静脉给予西妥昔单抗 pcpl INPs 可显著抑制肿瘤生长并提高存活率。通过 LC-MS/MS 在荷瘤小鼠中评估了前药和母体药物的药代动力学和器官分布。在肿瘤组织中未发现纳米颗粒或 INPs 的总蓄积增加。然而，对于包封的前药，观察到持续的 pcpl 水平和持续的紫杉醇转化和释放，这可能表明形成了药物库。总体结果表明，这种有前途的靶向平台具有改善肺癌和其他 EGFR 阳性肿瘤治疗的潜力。
