Epidermal growth factor receptor (EGFR) is overexpressed in many cancers and therefore serves as an excellent target for prodrug activation. Functionalised trans-cyclooctenes (TCO) were conjugated to an EGFR antibody (cetuximab), providing a reagent for pre-targeting and localisation of the bioorthogonal reagent. The TCOs react with a 4-azidobenzyl carbamate doxorubicin prodrug via a [3 + 2]-cycloaddition and subsequent self-immolation leads to release of doxorubicin (click-and-release). In vitro cell-based assays demonstrated proof-of-concept, that cetuximab conjugated to highly strained TCO (AB-d-TCO) could bind to the EGFR in a melanoma cell line, and selectively activate the doxorubicin prodrug. In a non-EGFR expressing melanoma cell line, no significant prodrug activation was observed. In vivo experiments using this combination of AB-d-TCO and the azido-doxorubicin prodrug in a murine melanoma model revealed no significant anti-tumour activity or increased survival, suggesting there was insufficient prodrug activation and drug release at the tumour site.

表皮生长因子受体 (EGFR) 在许多癌症中过度表达，因此可作为前药激活的极好靶点。功能化的反式环辛烯 (TCO) 与 EGFR 抗体（西妥昔单抗）结合，为生物正交试剂的预靶向和定位提供了一种试剂。TCO 与 4-叠氮苄基氨基甲酸酯多柔比星前药通过 [3 + 2]-环加成反应，随后的自焚导致多柔比星的释放（点击释放）。体外基于细胞的检测证明了概念验证，即与高度紧张的 TCO (AB-d-TCO) 结合的西妥昔单抗可以与黑色素瘤细胞系中的 EGFR 结合，并选择性地激活多柔比星前药。在不表达EGFR的黑色素瘤细胞系中，未观察到显着的前药活化。在小鼠黑色素瘤模型中使用 AB-d-TCO 和叠氮基多柔比星前药的这种组合进行的体内实验显示，没有显着的抗肿瘤活性或存活率增加，表明肿瘤部位的前药激活和药物释放不足。