Monomeric or Homodimer Conjugates of Fibroblast Activation Protein Inhibitor and Cyanine 7 Bearing a Meso-Chloride as Near-Infrared Fluorescence Probes: Design, Synthesis, and Comparative In Vivo Imaging of Distinct Breast Cancer Subtypes

Abstract

Intraoperative fluorescence navigation can illuminate the tumor, directing surgeons to accurately achieve negative margins, which not only reduces recurrence but also minimizes the incidence of complications. Herein, we developed two near-infrared fluorescent probes FAPI-Cy7-Cl (Em_max = 820 nm) and (FAPI)₂-Cy7-Cl (Em_max = 823 nm) with prolonged tumor retention (>72 h) and high target-to-background ratios (up to 4.5) based on the conjugation of pan-cancer targeted fibroblast activation protein inhibitor (FAPI) and the “tumor-seeking” Cyanine 7 bearing a meso-chloride and a cyclohexenyl skeleton (Cy7-Cl). Specifically, FAPI-Cy7-Cl exhibited superior imaging performance in both estrogen receptor α positive breast cancer (MCF-7) and triple-negative breast cancer (TNBC) (MDA-MB-231) subtypes in mouse models. Notably, in the MDA-MB-231 tumor-bearing model, the tumor-to-liver ratio (T/L) of FAPI-Cy7-Cl increased rapidly after 2 h postinjection, reaching nearly 4.5 at 48 h, making it an optimal imaging probe for guiding TNBC surgery resection.