LAUSR

Simple Summary One of the goals of nanoplatform-based cancer therapy is to achieve tumor accumulation of anticancer agents. We have focused on PIP-based KRAS inhibitor KR12 (pyrrole–imidazole polyamide indole-seco–CBI conjugate), which has been reported to exhibit tumor growth inhibition in a xenograft mouse model. To evaluate the tumor accumulation property of KR12, we have synthesized a fluorescently labeled KR12 derivative (KR12-TAMRA) and employed a chicken egg tumor assay, a simple and versatile assay to examine tumor accumulation. Our results show that KR12-TAMRA accumulates specifically in the tumor when injected into a fertilized chicken egg transplanted with human cancer cells. We also demonstrate nuclear accumulation of KR12-TAMRA. Finally, inhibition of tumor growth in the chorioallantoic membrane (CAM) assay is shown. These results uncover a number of attractive features of PIP-based KR12 for cancer therapy. Abstract Background: The KRAS inhibitor KR12, based on pyrrole-imidazole polyamide (PIP), has been developed and shown to exhibit efficacy in mouse experiments. Because some PIP species exhibit tumor accumulation capability, we decided to evaluate whether the PIP portion of KR12 exhibits tumor accumulation. We employed the CAM assay that provides a simple method for tumor accumulation evaluation. Methods: KR12 PIP was synthesized and conjugated to TAMRA to produce a fluorescently labeled reagent (KR12-TAMRA). This reagent was injected into a fertilized chicken egg that has been transplanted with human cancer cells. Distribution of the red fluorescence was examined by cutting out tumor as well as various organs from the embryo. Results: The red fluorescence of KR12-TAMRA was found to overlap with the green fluorescence of the tumor formed with GFP-expressing cancer cells. We also observed nuclear localization of KR12-TAMRA. Treatment of KR12 that contained the alkylating agent CBI in the tumor-bearing chicken egg resulted in tumor growth inhibition. Conclusions: KR12 contains a PIP that has two key features: tumor accumulation and nuclear localization. KR12 conjugated with CBI exhibits inhibition of tumor growth in the CAM model.