{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T13:00:19Z","timestamp":1775134819679,"version":"3.50.1"},"reference-count":32,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T00:00:00Z","timestamp":1775001600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Purpose: Prostate-specific membrane antigen (PSMA) is a well-established molecular target in prostate cancer (PCa). Both radionuclide imaging and near-infrared fluorescence (NIRF) imaging offer high sensitivity for in vivo tumor detection. PSMA-targeted dual-modality probes integrating these two imaging techniques provide complementary preoperative and intraoperative tumor visualization, thereby improving surgical guidance in PCa. In this study, we aimed to develop a novel dual-labeled PSMA probe combining radioactive and fluorescent properties to achieve precise tumor delineation during radical prostatectomy (RP). Methods: A high-affinity PSMA-targeted fluorescent probe (PSMA-DF) was synthesized using solid-phase synthesis. Subsequent radiolabeling with the radionuclide [68Ga]Ga yielded the successful generation of a dual-modal PSMA-targeted molecular probe, namely [68Ga]Ga-PSMA-DF. The probe was systematically evaluated both in vitro and in vivo, and its safety profile was assessed through acute toxicity testing. Tumor-bearing nude mouse models were established using PSMA-positive 22Rv1 and PSMA-negative PC-3 PCa cell lines. Imaging performance, tumor-targeting specificity, and biodistribution of the probe were comprehensively evaluated using micro-PET imaging, in vivo fluorescence imaging, and biodistribution studies. Results: High-quality and high-purity PSMA-DF was successfully prepared, which exhibited excellent optical properties. Following radiolabeling with [68Ga]Ga, a dual-modality radionuclide-fluorescence probe ([68Ga]Ga-PSMA-DF) was successfully constructed. In vitro cellular uptake studies demonstrated that 22Rv1 cells had relatively high uptake of the probe, reaching 7.34 \u00b1 0.55 IA%\/106 cells at 120 min. In contrast, PC-3 cells and blocked 22Rv1 cells displayed minimal uptake, confirming the specific targeting ability of the probe. In vivo evaluations were conducted on tumor-bearing mice using micro-PET\/CT and NIRF imaging. The results revealed that [68Ga]Ga-PSMA-DF achieved high specific tumor accumulation in 22Rv1 xenografts, with the peak tumor uptake (SUVmax = 1.748 \u00b1 0.132) and tumor-to-muscle ratio (11.542 \u00b1 1.511) observed at 120 min. Notably, high-contrast fluorescence imaging was also achieved at later time points, yielding a tumor-to-background ratio (TBR) of 6.559 \u00b1 1.415 at 48 h. Notably, ex vivo biodistribution data were consistent with in vivo imaging findings. Conclusions: This preclinical study demonstrates that [68Ga]Ga-PSMA-DF exhibits high and specific uptake in PCa models, supporting its potential as a dual-modality tracer for both PET\/CT imaging and real-time intraoperative fluorescence guidance during PCa surgery.<\/jats:p>","DOI":"10.3390\/ph19040564","type":"journal-article","created":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T11:51:03Z","timestamp":1775130663000},"page":"564","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Preclinical Study of a PSMA Ligand-Based Dual-Modality Probe for Radical Prostatectomy"],"prefix":"10.3390","volume":"19","author":[{"given":"Haoxi","family":"Zhou","sequence":"first","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"},{"name":"Chinese PLA Medical School, Chinese PLA General Hospital, Beijing 100853, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhiqiang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"},{"name":"Chinese PLA Medical School, Chinese PLA General Hospital, Beijing 100853, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Long","family":"Yi","sequence":"additional","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baojun","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaoxi","family":"Niu","sequence":"additional","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6908-6768","authenticated-orcid":false,"given":"Xu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Urology, The Third Medical Center, Chinese PLA General Hospital, Beijing 100089, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,1]]},"reference":[{"key":"ref_1","first-page":"12","article-title":"Cancer statistics, 2024","volume":"74","author":"Siegel","year":"2024","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1159\/000513258","article-title":"Localized and Locally Advanced Prostate Cancer: Treatment Options","volume":"99","author":"Achard","year":"2021","journal-title":"Oncology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1016\/S1470-2045(18)30357-7","article-title":"Robot-assisted laparoscopic prostatectomy versus open radical retropubic prostatectomy: 24-month outcomes from a randomised controlled study","volume":"19","author":"Coughlin","year":"2018","journal-title":"Lancet Oncol."},{"key":"ref_4","first-page":"1.e17","article-title":"Positive surgical margins at radical prostatectomy in the United States: Institutional variations and predictive factors","volume":"38","author":"Pooli","year":"2020","journal-title":"Urol. Oncol. Semin. Orig. Investig."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/s11701-019-00954-x","article-title":"Risk factors of positive surgical margins after robot-assisted radical prostatectomy in high-volume center: Results in 732 cases","volume":"14","author":"Porcaro","year":"2020","journal-title":"J. Robot. Surg."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ullah, Z., Roy, S., Gu, J., Soe, S.K., Jin, J., and Guo, B. (2024). NIR-II Fluorescent Probes for Fluorescence-Imaging-Guided Tumor Surgery. Biosensors, 14.","DOI":"10.3390\/bios14060282"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1038\/s41551-019-0494-0","article-title":"First-in-human liver-tumour surgery guided by multispectral fluorescence imaging in the visible and near-infrared-I\/II windows","volume":"4","author":"Hu","year":"2020","journal-title":"Nat. Biomed. Eng."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.1038\/nm.2472","article-title":"Intraoperative tumor-specific fluorescence imaging in ovarian cancer by folate receptor-\u03b1 targeting: First in-human results","volume":"17","author":"Themelis","year":"2011","journal-title":"Nat. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1002\/pros.21413","article-title":"Imaging prostate cancer lymph node metastases with a multimodality contrast agent","volume":"72","author":"Hall","year":"2012","journal-title":"Prostate"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/s12253-008-9104-2","article-title":"Heterogeneity of prostate-specific membrane antigen (PSMA) expression in prostate carcinoma with distant metastasis","volume":"15","author":"Mannweiler","year":"2009","journal-title":"Pathol. Oncol. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"596","DOI":"10.2967\/jnumed.120.257238","article-title":"Current and emerging clinical applications of PSMA PET diagnostic imaging for prostate cancer","volume":"62","author":"Farolfi","year":"2021","journal-title":"J. Nucl. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6824","DOI":"10.7150\/thno.36739","article-title":"PSMA-targeting agents for radio- and fluorescence-guided prostate cancer surgery","volume":"9","author":"Derks","year":"2019","journal-title":"Theranostics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1007\/s00259-016-3490-6","article-title":"Comparison of (68)Ga-labelled PSMA-11 and (11)C-choline in the detection of prostate cancer metastases by PET\/CT","volume":"44","author":"Schwenck","year":"2017","journal-title":"Eur. J. Nucl. Med. Mol. Imaging"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"17S","DOI":"10.2967\/jnumed.116.186775","article-title":"Glu-Ureido\u2013Based Inhibitors of Prostate-Specific Membrane Antigen: Lessons Learned During the Development of a Novel Class of Low-Molecular-Weight Theranostic Radiotracers","volume":"58","author":"Kopka","year":"2017","journal-title":"J. Nucl. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2774","DOI":"10.1007\/s00259-022-05775-z","article-title":"Synthesis, preclinical evaluation, and first-in-human study of Al18F-PSMA-Q for prostate cancer imaging","volume":"49","author":"Wu","year":"2022","journal-title":"Eur. J. Nucl. Med. Mol. Imaging"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"234","DOI":"10.2967\/jnumed.119.233064","article-title":"Hybrid Tracers Based on Cyanine Backbones Targeting Prostate-Specific Membrane Antigen: Tuning Pharmacokinetic Properties and Exploring Dye\u2013Protein Interaction","volume":"61","author":"Hensbergen","year":"2020","journal-title":"J. Nucl. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"71","DOI":"10.2967\/jnumed.118.212720","article-title":"Synthesis and Preclinical Characterization of the PSMA-Targeted Hybrid Tracer PSMA-I&F for Nuclear and Fluorescence Imaging of Prostate Cancer","volume":"60","author":"Schottelius","year":"2019","journal-title":"J. Nucl. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"79S","DOI":"10.2967\/jnumed.115.170720","article-title":"The Rise of PSMA Ligands for Diagnosis and Therapy of Prostate Cancer","volume":"57","author":"Babich","year":"2016","journal-title":"J. Nucl. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1056\/NEJMoa2107322","article-title":"Lutetium-177\u2013PSMA-617 for Metastatic Castration-Resistant Prostate Cancer","volume":"385","author":"Sartor","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1021\/acs.chemrev.1c00553","article-title":"Versatile types of inorganic\/organic NIR-IIa\/IIb fluorophores: From strategic design toward molecular imaging and theranostics","volume":"122","author":"Liu","year":"2022","journal-title":"Chem. Rev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1038\/s41551-021-00773-2","article-title":"A phosphorescent probe for in vivo imaging in the second near-infrared window","volume":"6","author":"Chang","year":"2022","journal-title":"Nat. Biomed. Eng."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"17168","DOI":"10.1039\/D1NR04625J","article-title":"Near-infrared light-responsive hybrid hydrogels for the synergistic chemo-photothermal therapy of oral cancer","volume":"13","author":"Wu","year":"2021","journal-title":"Nanoscale"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1007\/s10147-018-1365-5","article-title":"The effects of intraoperative ICG fluorescence angiography in laparoscopic low anterior resection: A propensity score-matched study","volume":"24","author":"Wada","year":"2019","journal-title":"Int. J. Clin. Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3073","DOI":"10.1007\/s13277-013-1399-2","article-title":"Breast cancer sentinel lymph node mapping using near-infrared guided indocyanine green in comparison with blue dye","volume":"35","author":"Guo","year":"2014","journal-title":"Tumor Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1162","DOI":"10.1016\/j.eururo.2013.11.017","article-title":"Fluorescence-enhanced robotic radical prostatectomy using real-time lymphangiography and tissue marking with percutaneous injection of unconjugated indocyanine green: The initial clinical experience in 50 patients","volume":"65","author":"Manny","year":"2014","journal-title":"Eur. Urol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1158\/1078-0432.CCR-18-0803","article-title":"Evaluation of Novel Prostate-Specific Membrane Antigen-Targeted Near-Infrared Imaging Agent for Fluorescence-Guided Surgery of Prostate Cancer","volume":"25","author":"Kularatne","year":"2019","journal-title":"Clin. Cancer Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"16645","DOI":"10.1021\/acs.jmedchem.5c01480","article-title":"Synthesis and Preclinical Evaluation of Dual-Mode Fluorescent 18F-PET Tracers Targeting PSMA","volume":"68","author":"Lozada","year":"2025","journal-title":"J. Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"639","DOI":"10.2967\/jnumed.117.201293","article-title":"PSMA-11\u2013Derived Dual-Labeled PSMA Inhibitors for Preoperative PET Imaging and Precise Fluorescence-Guided Surgery of Prostate Cancer","volume":"59","author":"Baranski","year":"2018","journal-title":"J. Nucl. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.eururo.2025.03.010","article-title":"First-in-human Study of a Dual-modality Prostate-specific Membrane Antigen-targeted Probe for Preoperative Positron Emission Tomography\/Computed Tomography Imaging and Intraoperative Fluorescence Imaging in Prostate Cancer","volume":"87","author":"Chen","year":"2025","journal-title":"Eur. Urol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.2967\/jnumed.118.221671","article-title":"Preclinical Evaluation and Pilot Clinical Study of Al18F-PSMA-BCH for Prostate Cancer PET Imaging","volume":"60","author":"Liu","year":"2019","journal-title":"J. Nucl. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2044","DOI":"10.1021\/acs.molpharmaceut.0c00199","article-title":"Combined Application of Albumin-Binding [177Lu]Lu-PSMA-ALB-56 and Fast-Cleared PSMA Inhibitors: Optimization of the Pharmacokinetics","volume":"17","author":"Borgna","year":"2020","journal-title":"Mol. Pharm."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1744806917697008","DOI":"10.1177\/1744806917697008","article-title":"A role for the locus coeruleus in the analgesic efficacy of N-acetylaspartylglutamate peptidase (GCPII) inhibitors ZJ43 and 2-PMPA","volume":"13","author":"Nonaka","year":"2017","journal-title":"Mol. Pain"}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/4\/564\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T11:59:59Z","timestamp":1775131199000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/4\/564"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,1]]},"references-count":32,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["ph19040564"],"URL":"https:\/\/doi.org\/10.3390\/ph19040564","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,1]]}}}