Inhibition of epidermal growth factor receptor-mediated signaling by "Combi-triazene" BJ2000, a new probe for Combi-Targeting postulates.

摘要: The Combi-Targeting concept postulates that a molecule termed combi-molecule (C-molecule) with binary epidermal growth factor receptor (EGFR) targeting/DNA-damaging properties and the ability to be hydrolyzed another EGFR inhibitor should induce sustained antiproliferative activity in cells overexpressing EGFR. Because we postulate affinity of C-molecule its hydrolytic metabolites are critical parameters for potency against EGFR-overexpressing cells, synthesized BJ2000 (IC50 = 0.1 μM, competitive binding at ATP site), novel can decompose into 6-amino-4-anilinoquinazoline FD105 0.2 μM). Studies using A431 revealed could damage DNA block factor-stimulated autophosphorylation by partially irreversible mechanism. Blockade subsequently induced inhibition mitogen-activated protein kinase activation c-fos gene expression. Enzyme-linked immunosorbent assay factor-mediated stimulation proliferation assays EGFR-expressing NIH3T3HER14 demonstrated preferential EGFR-targeting BJ2000, more importantly suggest blockade phosphorylation this drug translate significant inhibitory effects. These culminated effects as confirmed sulforhodamine B assay. Five days after 2-h treatment, retained effect whereas reversible metabolite almost completely lost activity. This result toto lend support according which molecular conjugate kept small enough interact designed degrade same target plus DNA-damaging species may cells.