Presently, surgical resection offers the only chance of cure for patients with pancreatic adenocarcinoma. While the goal of resection is to obtain R0 (negative) margins, the reported R1 (positive tumor within 1 mm) margin resection rates of patients undergoing pancreatic resection range up to 75% resulting in an unmet need to intraoperatively identify live PDAC tumor. The extent of vascular involvement of tumor, potential surrounding inflammation, desmoplastic reaction, and dense tissue abutting these critical vessels may impact successful resections in potentially patients with borderline resectable tumors. While recent advances in fluorescent-guided surgery and on-going active clinical trials that utilize NIR-fluorescent labeled monoclonal antibodies to target PDAC have demonstrated PDAC tumor enhancement in some cases where tumors were positive/upregulated expression of the receptor, these trials also highlight the need for additional molecular probes to address biomarker heterogeneity in pancreatic cancer. These NIR-fluorescent imaging trials also highlight the need to overcome potential fluorescent signal quenching by blood and adjacent blood vessels to improve resection accuracy. To address these limitations, we have investigated extracellular acidic pH as a biomarker for pancreatic cancer and conducted a Phase I safety assessment of intraoperative Multispectral Optoacoustic Tomography (IIT trial NCT04365413). Clinical MSOT study indicated safety of the MSOT device in a human surgical setting and our murine in vivo results suggest that tumor-specific contrast agents improve tumor identification, our objective to develop VD-ZW800C contrast agent to facilitate image-guided surgery of PDAC. We have developed a second generation optoacoustically detectable pancreatic cancer specific probe, VD- ZW800C to facilitate MSOT- or NIR-fluorescent- guided resection of pancreatic cancer. The overarching hypothesis is that VD-ZW800C will have an acceptable safety profile and use of VD-ZW800C will improve the separation of PDAC tumor tissue from benign or fibrosis (resulting from chemo- or radiation therapy) tissues in real-time using clinical MSOT or NIR fluorescent imaging with greater specificity than frozen pathology. Based upon this hypothesis, we propose the following objectives: 1) Develop VD-ZW800C that is detectable using MSOT and NIR fluorescence; 2) Evaluate preclinical toxicology and pharmacokinetics; and 3) Assess optimized VD-ZW800C to facilitate detection of PDAC in Oncopig using MSOT and NIR-guided surgery and assess VD- ZW800C microdose 1st-in human. Successful completion of these specific aims will produce VD-ZW800C as a pancreatic cancer-specific probe detectable by MSOT or NIR fluorescence. We will test our VD-ZW800C probe as both an MSOT and NIR-fluorescent detectable imaging probe in the Oncopig and 1st in human. Our long-term impact will be in demonstrating the feasibility of VD-ZW800C as a PDAC specific targeted image guided surgica...