{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T22:14:45Z","timestamp":1788300885393,"version":"build-2803163510"},"reference-count":50,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2015,8,26]],"date-time":"2015-08-26T00:00:00Z","timestamp":1440547200000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["R01CA140617"],"award-info":[{"award-number":["R01CA140617"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,10,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Prostate cancer (PCa) is the second most common cause of cancer death among American men after lung cancer. Unfortunately, current therapies do not provide effective treatments for patients with advanced, metastatic, or hormone refractory disease. Therefore, we seek to generate therapeutic agents for a novel PCa treatment strategy by delivering a suicide enzyme (yCDtriple) to a cell membrane bound biomarker found on PCa cells (prostate-specific membrane antigen (PSMA)). This approach has resulted in a new PCa treatment strategy reported here as inhibitor-directed enzyme prodrug therapy (IDEPT). The therapeutic agents described were generated using a click chemistry reaction between the unnatural amino acid (p-azidophenylalanine (pAzF)) incorporated into yCDtriple and the dibenzylcyclooctyne moiety of our PSMA targeting agent (DBCO-PEG4-AH2-TG97). After characterization of the therapeutic agents, we demonstrate significant PCa cell killing of PSMA-positive cells. Importantly, we demonstrate that this click chemistry approach can be used to efficiently couple a therapeutic protein to a targeting agent and may be applicable to the ablation of other types of cancers and\/or malignancies.<\/jats:p>","DOI":"10.1021\/bc500362n","type":"journal-article","created":{"date-parts":[[2014,8,26]],"date-time":"2014-08-26T13:18:26Z","timestamp":1409059106000},"page":"1752-1760","source":"Crossref","is-referenced-by-count":14,"title":["Development of Inhibitor-Directed Enzyme Prodrug Therapy\n(IDEPT) for Prostate Cancer"],"prefix":"10.1021","volume":"25","author":[{"given":"Stacy E.","family":"Martin","sequence":"first","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanushree","family":"Ganguly","sequence":"additional","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerhard R.","family":"Munske","sequence":"additional","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melody D.","family":"Fulton","sequence":"additional","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark R.","family":"Hopkins","sequence":"additional","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clifford E.","family":"Berkman","sequence":"additional","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margaret E.","family":"Black","sequence":"additional","affiliation":[{"name":"Washington State University , , , ,","place":["Pullman, Washington, United States, 99164-7520"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2014,8,26]]},"reference":[{"key":"2026090117322605900_cit1","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1146\/annurev.biophys.35.101105.121507","article-title":"Expanding the genetic code","volume":"35","author":"Wang","year":"2006","journal-title":"Annu.\nRev. 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Ed."},{"key":"2026090117322605900_cit4","doi-asserted-by":"crossref","first-page":"6571","DOI":"10.1021\/bi9007726","article-title":"Click chemistry\nand bioorthogonal reactions: unprecedented selectivity in the labeling\nof biological molecules","volume":"48","author":"Best","year":"2009","journal-title":"Biochemistry"},{"key":"2026090117322605900_cit5","doi-asserted-by":"crossref","first-page":"3878","DOI":"10.1002\/anie.201008178","article-title":"Genetically\nencoded copper-free click chemistry","volume":"50","author":"Plass","year":"2011","journal-title":"Angew.\nChem., Int. Ed."},{"key":"2026090117322605900_cit6","doi-asserted-by":"crossref","first-page":"9918","DOI":"10.1021\/ja303904e","article-title":"Synthesis of bispecific\nantibodies using genetically encoded unnatural amino acids","volume":"134","author":"Kim","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"2026090117322605900_cit7","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1021\/jo026615r","article-title":"Click chemistry to construct fluorescent\noligonucleotides for DNA sequencing","volume":"68","author":"Seo","year":"2002","journal-title":"J. Org.\nChem."},{"key":"2026090117322605900_cit8","doi-asserted-by":"crossref","first-page":"11486","DOI":"10.1021\/ja803086r","article-title":"Second-generation\ndifluorinated cyclooctynes for copper-free click chemistry","volume":"130","author":"Codelli","year":"2008","journal-title":"J. Am. Chem. Soc."},{"key":"2026090117322605900_cit9","doi-asserted-by":"crossref","first-page":"1570","DOI":"10.1021\/bc800077y","article-title":"In vivo tumor cell targeting with\n\u201cclick\u201d nanoparticles","volume":"19","author":"von Maltzahn","year":"2008","journal-title":"Bioconjugate\nChem."},{"key":"2026090117322605900_cit10","doi-asserted-by":"crossref","first-page":"15046","DOI":"10.1021\/ja044996f","article-title":"A strain-promoted\n[3 + 2] azide-alkyne\ncycloaddition for covalent modification of biomolecules in living\nsystems","volume":"126","author":"Agard","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"2026090117322605900_cit11","first-page":"1313","article-title":"Purification\nand some properties of cytosine deaminase from bakers\u2019 yeast","volume":"53","author":"Katsuragi","year":"1989","journal-title":"Agric. Biol. Chem."},{"key":"2026090117322605900_cit12","first-page":"6649","article-title":"Yeast cytosine deaminase\nimproves radiosensitization and bystander effect by 5-fluorocytosine\nof human colorectal cancer xenografts","volume":"60","author":"Kievit","year":"2000","journal-title":"Cancer\nRes."},{"key":"2026090117322605900_cit13","doi-asserted-by":"crossref","first-page":"8302","DOI":"10.1073\/pnas.91.17.8302","article-title":"Metabolism\nof 5-fluorocytosine to 5-fluorouracil in human colorectal tumor cells\ntransduced with the cytosine deaminase gene: significant antitumor\neffects when only a small percentage of tumor cells express cytosine\ndeaminase","volume":"91","author":"Huber","year":"1994","journal-title":"Proc. Natl. Acad. Sci. 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Pharmaceutics"},{"key":"2026090117322605900_cit16","doi-asserted-by":"crossref","first-page":"3168","DOI":"10.1158\/1078-0432.CCR-05-2729","article-title":"Novel cytosine deaminase fusion gene\nenhances the effect of radiation on breast cancer in bone by reducing\ntumor burden, osteolysis, and skeletal fracture","volume":"12","author":"Goblirsch","year":"2006","journal-title":"Clin. Cancer Res."},{"key":"2026090117322605900_cit17","first-page":"1417","article-title":"Superiority of yeast over bacterial\ncytosine deaminase for enzyme\/prodrug gene therapy in colon cancer\nxenografts","volume":"59","author":"Kievit","year":"1999","journal-title":"Cancer Res."},{"key":"2026090117322605900_cit18","first-page":"2719","article-title":"Intratumoral generation of 5-fluorouracil\nmediated\nby an antibody-cytosine deaminase conjugate in combination with 5-fluorocytosine","volume":"54","author":"Wallace","year":"1994","journal-title":"Cancer Res."},{"key":"2026090117322605900_cit19","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1038\/mt.sj.6300120","article-title":"Phase I trial of replication-competent\nadenovirus-mediated suicide gene therapy combined with IMRT for prostate\ncancer","volume":"15","author":"Freytag","year":"2007","journal-title":"Mol. Ther."},{"key":"2026090117322605900_cit20","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1126\/science.1107387","article-title":"Computational\nthermostabilization\nof an enzyme","volume":"308","author":"Korkegian","year":"2005","journal-title":"Science"},{"key":"2026090117322605900_cit21","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1016\/j.jmb.2008.01.002","article-title":"Yeast cytosine deaminase\nmutants\nwith increased thermostability impart sensitivity to 5-fluorocytosine","volume":"377","author":"Stolworthy","year":"2008","journal-title":"J. Mol. Biol."},{"key":"2026090117322605900_cit22","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1093\/neuonc\/nor199","article-title":"Brain tumor\neradication and prolonged survival from intratumoral conversion of\n5-fluorocytosine to 5-fluorouracil using a nonlytic retroviral replicating\nvector","volume":"14","author":"Ostertag","year":"2012","journal-title":"Neuro-Oncology"},{"key":"2026090117322605900_cit23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s002940050169","article-title":"Characterization of\nthe saccharomyces\ncerevisiae FCY1 gene encoding cytosine deaminase and its homologue\nFCA1 of candida albicans","volume":"31","author":"Erbs","year":"1997","journal-title":"Curr. Genet."},{"key":"2026090117322605900_cit24","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1006\/prep.1997.0839","article-title":"Cloning, overexpression, and purification\nof cytosine\ndeaminase from saccharomyces cerevisiae","volume":"12","author":"Hayden","year":"1998","journal-title":"Protein\nExpression Purif."},{"key":"2026090117322605900_cit25","doi-asserted-by":"crossref","first-page":"2256","DOI":"10.1002\/(SICI)1097-0142(19980601)82:11<2256::AID-CNCR22>3.0.CO;2-S","article-title":"Prostate\nspecific membrane antigen expression in prostatic intraepithelial\nneoplasia and adenocarcinoma","volume":"82","author":"Bostwick","year":"1998","journal-title":"Cancer"},{"key":"2026090117322605900_cit26","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1002\/(SICI)1097-0045(20000301)42:4<318::AID-PROS10>3.0.CO;2-L","article-title":"Serum levels of PSMA","volume":"42","author":"Murphy","year":"2000","journal-title":"Prostate"},{"key":"2026090117322605900_cit27","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/S0090-4295(96)00184-7","article-title":"Upregulation of prostate-specific\nmembrane antigen after androgen-deprivation therapy","volume":"48","author":"Wright","year":"1996","journal-title":"Urology"},{"key":"2026090117322605900_cit28","first-page":"3192","article-title":"Five different anti-prostate-specific membrane antigen\n(PSMA) antibodies confirm PSMA expression in tumor-associated neovasculature","volume":"59","author":"Chang","year":"1999","journal-title":"Cancer Res."},{"key":"2026090117322605900_cit29","first-page":"2674","article-title":"Prostate-specific membrane antigen is produced in tumor-associated\nneovasculature","volume":"5","author":"Chang","year":"1999","journal-title":"Clin. Cancer Res."},{"key":"2026090117322605900_cit30","doi-asserted-by":"crossref","first-page":"4504","DOI":"10.1021\/jm800111u","article-title":"Synthesis and evaluation\nof technetium-99m-and rhenium-labeled\ninhibitors of the prostate-specific membrane antigen (PSMA)","volume":"51","author":"Banerjee","year":"2008","journal-title":"J. Med. Chem."},{"key":"2026090117322605900_cit31","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1002\/cmdc.200500044","article-title":"Design\nand synthesis of a PSMA inhibitor-doxorubicin conjugate for targeted\nprostate cancer therapy","volume":"1","author":"Jayaprakash","year":"2006","journal-title":"ChemMedChem"},{"key":"2026090117322605900_cit32","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1021\/mp900069d","article-title":"Prostate-specific\nmembrane antigen targeted imaging and therapy of prostate cancer using\na PSMA inhibitor as a homing ligand","volume":"6","author":"Kularatne","year":"2009","journal-title":"Mol. Pharmaceutics"},{"key":"2026090117322605900_cit33","doi-asserted-by":"crossref","first-page":"2042","DOI":"10.2967\/jnumed.109.066589","article-title":"Assessment\nof an 18F-labeled phosphoramidate peptidomimetic as a new prostate-specific\nmembrane antigen-targeted imaging agent for prostate cancer","volume":"50","author":"Lapi","year":"2009","journal-title":"J. Nucl. Med."},{"key":"2026090117322605900_cit34","doi-asserted-by":"crossref","first-page":"7767","DOI":"10.1021\/jm100729b","article-title":"Synthesis and biological analysis\nof prostate-specific membrane antigen-targeted anticancer prodrugs","volume":"53","author":"Kularatne","year":"2010","journal-title":"J. Med. Chem."},{"key":"2026090117322605900_cit35","doi-asserted-by":"crossref","first-page":"12658","DOI":"10.1021\/bi801883v","article-title":"Pseudoirreversible\ninhibition of prostate-specific membrane antigen by phosphoramidate\npeptidomimetics","volume":"47","author":"Liu","year":"2008","journal-title":"Biochemistry"},{"key":"2026090117322605900_cit36","doi-asserted-by":"crossref","first-page":"3931","DOI":"10.1016\/j.bmcl.2012.04.110","article-title":"Targeting prostate cancer cells with\na multivalent PSMA inhibitor-guided streptavidin conjugate","volume":"22","author":"Liu","year":"2012","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"2026090117322605900_cit37","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1002\/pros.20753","article-title":"Cell-surface labeling\nand internalization by a fluorescent inhibitor of prostate-specific\nmembrane antigen","volume":"68","author":"Liu","year":"2008","journal-title":"Prostate"},{"key":"2026090117322605900_cit38","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1002\/bit.22070","article-title":"High-level cell-free synthesis yields\nof proteins containing\nsite-specific non-natural amino acids","volume":"102","author":"Goerke","year":"2009","journal-title":"Biotechnol.\nBioeng."},{"key":"2026090117322605900_cit39","doi-asserted-by":"crossref","first-page":"9026","DOI":"10.1021\/ja027007w","article-title":"Addition of p-azido-l-phenylalanine\nto the genetic\ncode of escherichia coli","volume":"124","author":"Chin","year":"2002","journal-title":"J. Am. Chem. Soc."},{"key":"2026090117322605900_cit40","doi-asserted-by":"crossref","first-page":"4270","DOI":"10.1021\/bi00799a018","article-title":"Baker\u2019s yeast\ncytosine deaminase. Some enzymatic\nproperties and allosteric inhibition by nucleosides and nucleotides","volume":"10","author":"Ipata","year":"1971","journal-title":"Biochemistry"},{"key":"2026090117322605900_cit41","doi-asserted-by":"crossref","first-page":"8957","DOI":"10.1021\/bi049720z","article-title":"Alanine-scanning\nmutagenesis reveals a cytosine deaminase mutant with altered substrate\npreference","volume":"43","author":"Mahan","year":"2004","journal-title":"Biochemistry"},{"key":"2026090117322605900_cit42","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/S0026-895X(25)13624-9","article-title":"A first step in the development of\ngene therapy for\ncolorectal carcinoma: cloning, sequencing, and expression of Escherichia\ncoli cytosine deaminase","volume":"43","author":"Austin","year":"1993","journal-title":"Mol. Pharmacol."},{"key":"2026090117322605900_cit43","doi-asserted-by":"crossref","first-page":"7013","DOI":"10.1016\/j.bmcl.2011.09.115","article-title":"Spacer length effects\non in vitro imaging and surface accessibility of fluorescent inhibitors\nof prostate specific membrane antigen","volume":"21","author":"Liu","year":"2011","journal-title":"Bioorg.\nMed. Chem. Lett."},{"key":"2026090117322605900_cit44","doi-asserted-by":"crossref","first-page":"11782","DOI":"10.1021\/ja0370037","article-title":"Adding amino acids with novel reactivity to the genetic\ncode of Saccharomyces cerevisiae","volume":"125","author":"Deiters","year":"2003","journal-title":"J. Am. Chem.\nSoc."},{"key":"2026090117322605900_cit45","doi-asserted-by":"crossref","first-page":"1521","DOI":"10.1016\/j.bmcl.2004.12.065","article-title":"In vivo incorporation\nof an alkyne into proteins in\nEscherichia coli","volume":"15","author":"Deiters","year":"2005","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"2026090117322605900_cit46","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.pep.2006.05.008","article-title":"Purification\nof prostate-specific\nmembrane antigen using conformational epitope-specific antibody-affinity\nchromatography","volume":"49","author":"Liu","year":"2006","journal-title":"Protein Expression Purif."},{"key":"2026090117322605900_cit47","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1093\/jac\/dkm389","article-title":"Antifungal serum\nconcentration monitoring: an update","volume":"61","author":"Goodwin","year":"2008","journal-title":"J. Antimicrob.\nChemother."},{"key":"2026090117322605900_cit48","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1128\/AAC.00705-08","article-title":"Antifungal\ntherapeutic drug monitoring:\nestablished and emerging indications","volume":"53","author":"Andes","year":"2009","journal-title":"Antimicrob.\nAgents Chemother."},{"key":"2026090117322605900_cit49","doi-asserted-by":"crossref","first-page":"4969","DOI":"10.1016\/j.bmc.2004.06.031","article-title":"Probing for a hydrophobic\na binding register in prostate-specific membrane antigen with phenylalkylphosphonamidates","volume":"12","author":"Maung","year":"2004","journal-title":"Bioorg. Med. Chem."},{"key":"2026090117322605900_cit50","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.bmc.2005.07.056","article-title":"Optimizing phenylethylphosphonamidates\nfor the inhibition of prostate-specific membrane antigen","volume":"14","author":"Wone","year":"2006","journal-title":"Bioorg. Med. Chem."}],"container-title":["Bioconjugate Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/bc500362n","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bcches\/article-pdf\/25\/10\/1752\/15672357\/bc500362n.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bcches\/article-pdf\/25\/10\/1752\/15672357\/bc500362n.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T21:32:38Z","timestamp":1788298358000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bcches\/article\/25\/10\/1752\/1475239\/Development-of-Inhibitor-Directed-Enzyme-Prodrug"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,8,26]]},"references-count":50,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2014,8,26]]},"published-print":{"date-parts":[[2014,10,15]]}},"URL":"https:\/\/doi.org\/10.1021\/bc500362n","relation":{},"ISSN":["1043-1802","1520-4812"],"issn-type":[{"value":"1043-1802","type":"print"},{"value":"1520-4812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,8,26]]}}}