NIR‐II Fluorescence Surgical Navigation for Gastric Cancer: Systematic Preclinical Validation Toward Clinical Application

ABSTRACT The precise intraoperative delineation of gastric tumors and their micrometastases remains a major challenge in surgical oncology. To address this, we developed a novel NIR‐II fluorescent probe named SPN01 targeting integrin αvβ3, a receptor upregulated in gastric cancer and associated tumor vasculature. SPN01 was constructed by conjugating RGD peptide to a high‐performance cyanine fluorophore with net neutral charge. In multiple preclinical models, SPN01 enabled high‐contrast visualization of sub‐centimeter orthotopic tumors, detection of occult peritoneal micrometastases as small as 0.5 mm, and specific identification of metastatic lymph nodes. Concurrently, near‐infrared II (NIR‐II, 900–1880 nm) fluorescence guidance facilitated the complete resection of tumors with clear margins, owing to the superior brightness of SPN01 in the NIR‐II window. Notably, ex vivo validation using human gastric cancer specimens confirmed target‐specific binding. Comprehensive pharmacokinetic and safety studies demonstrated rapid dual‐pathway clearance and no obvious toxicity. Following these findings, we conducted a first‐in‐human Phase I clinical trial involving 24 healthy volunteers, which demonstrated a favorable safety profile for SPN01. These findings support SPN01‐assisted NIR‐II fluorescence‐guided surgery as a promising candidate for clinical translation with the potential to enhance the completeness of gastric cancer resection.