This report describes the design, synthesis and evaluation of tumor-targeted polymer probes to visualize epidermal growth factor receptor (EGFR)-positive malignant tumors for successful resection via fluorescence guided endoscopic surgery. Fluorescent polymer probes of various molecular weights enabling passive accumulation in tumors via enhanced permeability and retention were prepared and evaluated, showing an optimal molecular weight of 200,000 g/mol for passive tumor targeting. Moreover, poly(N-(2-hydroxypropyl)methacrylamide)-based copolymers labeled with fluorescent dyes were targeted with the EGFR-binding oligopeptide GE-11 (YHWYGYTPQNVI), human EGF or anti-EGFR monoclonal antibody cetuximab were all able to actively target the surface of EGFR-positive tumor cells. Nanoprobes targeted with GE-11 and cetuximab showed the best targeting profile but differed in their tumor accumulation kinetics. Cetuximab increased tumor accumulation after 15 min, whereas GE 11 needed at least 4 h. Interestingly, after 4 h, there were no significant differences in tumor targeting, indicating the potential of oligopeptide targeting for fluorescence-navigated surgery. In conclusion, fluorescent polymer probes targeted by oligopeptide GE-11 or whole antibody are excellent tools for surgical navigation during oncological surgery of head and neck squamous cell carcinoma, due to their relatively simple design, synthesis and cost, as well as optimal pharmacokinetics and accumulation in tumors.

Center for Advanced Preclinical Imaging 1st Faculty of Medicine Charles University U Nemocnice 5 120 00 Prague 2 Czech Republic

Department of Otorhinolaryngology Head and Neck Surgery 1st Faculty of Medicine Charles University and University Hospital Motol 5 Úvalu 84 150 06 Prague 5 Czech Republic

Institute of Biophysics and Informatics 1st Faculty of Medicine Charles University Salmovská 1 120 00 Prague 2 Czech Republic

Institute of Macromolecular Chemistry Czech Academy of Sciences Heyrovského sq 2 162 06 Prague 6 Czech Republic

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