Prostate cancer is primarily fatal after it becomes metastatic and castration-resistant despite novel combined hormonal and chemotherapeutic regimens. Hence, new therapeutic concepts and drug delivery strategies are urgently needed for the eradication of this devastating disease. Here we report the highly specific, in situ click chemistry driven pretargeted delivery of cytotoxic drug carriers to PSMA(+) prostate cancer cells. Anti-PSMA 5D3 mAb and its F(ab')2 fragments were functionalized with trans-cyclooctene (TCO), labeled with a fluorophore, and used as pretargeting components. Human serum albumin (ALB) was loaded with the DM1 antitubulin agent, functionalized with PEGylated tetrazine (PEG4-Tz), labeled with a fluorophore, and used as the drug delivery component. The internalization kinetics of components and the therapeutic efficacy of the pretargeted click therapy were studied in PSMA(+) PC3-PIP and PSMA(-) PC3-Flu control cells. The F(ab')2 fragments were internalized faster than 5D3 mAb in PSMA(+) PC3-PIP cells. In the two-component pretargeted imaging study, both components were colocalized in a perinuclear location of the cytoplasm of PC3-PIP cells. Better colocalization was achieved when 5D3 mAb was used as the pretargeting component. Consecutively, the in vitro cell viability study shows a significantly higher therapeutic effect of click therapy in PC3-PIP cells when 5D3 mAb was used for pretargeting, compared to its F(ab')2 derivative. 5D3 mAb has a longer lifetime on the cell surface, when compared to its F(ab')2 analogue, enabling efficient cross-linking with the drug delivery component and increased efficacy. Pretargeting and drug delivery components were cross-linked via multiple bioorthogonal click chemistry reactions on the surface of PSMA(+) PC cells forming nanoclusters, which undergo fast cellular internalization and intracellular transport to perinuclear locations.

Department of Oncology The Sidney Kimmel Comprehensive Cancer Center The Johns Hopkins University School of Medicine 401 N Broadway Baltimore Maryland 21231 United States

Laboratory of Structural Biology Institute of Biotechnology of the Czech Academy of Sciences Prumyslova 595 Vestec 252 50 Czech Republic

The James Buchanan Brady Urologic Institute and Department of Urology Johns Hopkins School of Medicine 600 N Wolfe St Baltimore Maryland 21287 United States

The Russell H Morgan Department of Radiology and Radiological Science The Johns Hopkins University School of Medicine 720 Rutland Avenue Baltimore Maryland 21205 United States

Zobrazit více v PubMed

Reddy A; Roberts R; Shenoy D; Packianathan S; Giri S; Vijayakumar S Prostate Cancer Screening Guidelines for African American Veterans: A New Perspective. J. Natl. Med. Assoc 2018. PubMed

Ahlborg H; Nightingale AJ Mismatch between Scales of Knowledge in Nepalese Forestry: Epistemology, Power, and Policy Implications. Ecology and Society 2012, 17, 1.

Siegel RL; Miller KD; Jemal A Cancer Statistics, 2019. Ca-Cancer J. Clin 2019, 69, 7–34. PubMed

Fang JC; Faerber G; Samadder J Digital Rectal Examination for Prostate Cancer Screening Performed with Colonoscopy for Colon Cancer Screening: 2 for the Price of 1. Gastrointestinal Endoscopy 2017, 86, 1147–1150. PubMed

Zerbib M; Zelefsky MJ; Higano CS; Carroll PR Conventional Treatments of Localized Prostate Cancer. Urology 2008, 72, 25–35. PubMed

Frieling JS; Basanta D; Lynch CC Current and Emerging Therapies for Bone Metastatic Castration-Resistant Prostate Cancer. Cancer Control 2015, 22, 109–120. PubMed PMC

Gaya JM; Huguet J; Breda A; Palou J [Surgical Treatment of Local Disease in Metastatic Prostate Cancer.]. Arch Esp Urol 2018, 71, 288–297. PubMed

Vanneste BG; Van Limbergen EJ; van Lin EN; van Roermund JG; Lambin P Prostate Cancer Radiation Therapy: What Do Clinicians Have to Know? BioMed Res. Int 2016, 2016, 6829875. PubMed PMC

Das M Androgen Deprivation Therapy for Prostate Cancer. Lancet Oncol. 2017, 18, e567. PubMed

Sartor O; de Bono JS Metastatic Prostate Cancer. N. Engl. J. Med 2018, 378, 1653–1654. PubMed

Lee CH; Kantoff P Treatment of Metastatic Prostate Cancer in 2018. JAMA Oncol 2019, 5, 263. PubMed PMC

Osanto S; Van Poppel H Emerging Novel Therapies for Advanced Prostate Cancer. Ther. Adv. Urol 2012, 4, 3–12. PubMed PMC

Clarke JM; Armstrong AJ Novel Therapies for the Treatment of Advanced Prostate Cancer. Curr. Treat Options Oncol 2013, 14, 109–126. PubMed PMC

Poon DM; Ng J; Chan K Importance of Cycles of Chemotherapy and Postdocetaxel Novel Therapies in Metastatic Castration-Resistant Prostate Cancer. Prostate Int. 2015, 3, 51–55. PubMed PMC

Bouchelouche K; Choyke PL; Capala J Prostate Specific Membrane Antigen - a Target for Imaging and Therapy with Radionuclides. Discovery Medicine 2010, 9, 55–61. PubMed PMC

Bravaccini S; Puccetti M; Bocchini M; Ravaioli S; Celli M; Scarpi E; De Giorgi U; Tumedei MM; Raulli G; Cardinale L Psma Expression: A Potential Ally for the Pathologist in Prostate Cancer Diagnosis. Sci. Rep 2018, 8, 1. PubMed PMC

Mease RC; Foss CA; Pomper MG Pet Imaging in Prostate Cancer: Focus on Prostate-Specific Membrane Antigen. Curr. Top. Med. Chem 2013, 13, 951–962. PubMed PMC

Chatalic KLS; Veldhoven-Zweistra J; Bolkestein M; Hoeben S; Koning GA; Boerman OC; de Jong M; van Weerden WM A Novel in-111-Labeled Anti-Prostate-Specific Membrane Antigen Nanobody for Targeted Spect/Ct Imaging of Prostate Cancer. J. Nucl. Med 2015, 56, 1094–1099. PubMed

Chang SS Overview of Prostate-Specific Membrane Antigen. Rev. Urol 2004, 6 Suppl 10, S13–18. PubMed PMC

Langut Y; Talhami A; Mamidi S; Shir A; Zigler M; Joubran S; Sagalov A; Flashner-Abramson E; Edinger N; Klein S; et al. Psma-Targeted Polyinosine/Polycytosine Vector Induces Prostate Tumor Regression and Invokes an Antitumor Immune Response in Mice. Proc. Natl. Acad. Sci. U. S. A 2017, 114, 13655–13660. PubMed PMC

Wustemann T; Haberkorn U; Babich J; Mier W Targeting Prostate Cancer: Prostate-Specific Membrane Antigen Based Diagnosis and Therapy. Med. Res. Rev 2019, 39, 40–69. PubMed

Yao V; Berkman CE; Choi JK; O’Keefe DS; Bacich DJ Expression of Prostate-Specific Membrane Antigen (Psma), Increases Cell Folate Uptake and Proliferation and Suggests a Novel Role for Psma in the Uptake of the Non-Polyglutamated Folate, Folic Acid. Prostate 2009, 70, 305–316. PubMed

Mesters JR; Barinka C; Li W; Tsukamoto T; Majer P; Slusher BS; Konvalinka J; Hilgenfeld R Structure of Glutamate Carboxypeptidase Ii, a Drug Target in Neuronal Damage and Prostate Cancer. EMBO J. 2006, 25, 1375–1384. PubMed PMC

Barinka C; Rojas C; Slusher B; Pomper M Glutamate Carboxypeptidase Ii in Diagnosis and Treatment of Neurologic Disorders and Prostate Cancer. Curr. Med. Chem 2012, 19, 856–870. PubMed PMC

Pinto JT; Suffoletto BP; Berzin TM; Qiao CH; Lin S; Tong WP; May F; Mukherjee B; Heston WD Prostate-Specific Membrane Antigen: A Novel Folate Hydrolase in Human Prostatic Carcinoma Cells. Clin. Cancer Res 1996, 2, 1445–1451. PubMed

Carter RE; Feldman AR; Coyle JT Prostate-Specific Membrane Antigen Is a Hydrolase with Substrate and Pharmacologic Characteristics of a Neuropeptidase. Proc. Natl. Acad. Sci. U. S. A 1996, 93, 749–753. PubMed PMC

Rahbar K; Afshar-Oromieh A; Jadvar H; Ahmadzadehfar H Psma Theranostics: Current Status and Future Directions. Mol. Imaging 2018, 17, 153601211877606. PubMed PMC

Azad BB; Banerjee SR; Pullambhatla M; Lacerda S; Foss CA; Wang YC; Ivkov R; Pomper MG Evaluation of a Psma-Targeted Bnf Nanoparticle Construct. Nanoscale 2015, 7, 4432–4442. PubMed PMC

Chandran SS; Banerjee SR; Mease RC; Pomper MG; Denmeade SR Characterization of a Targeted Nanoparticle Functionalized with a Urea-Based Inhibitor of Prostate-Specific Membrane Antigen (Psma). Cancer Biol. Ther 2008, 7, 974–982. PubMed PMC

Jin W; Qin B; Chen ZJ; Liu H; Barve A; Cheng K Discovery of Psma-Specific Peptide Ligands for Targeted Drug Delivery. Int. J. Pharm 2016, 513, 138–147. PubMed

Liu TC; Nedrow-Byers JR; Hopkins MR; Wu LSY; Lee J; Reilly PTA; Berkman CE Targeting Prostate Cancer Cells with a Multivalent Psma Inhibitor-Guided Streptavidin Conjugate. Bioorg. Med. Chem. Lett 2012, 22, 3931–3934. PubMed PMC

Fuchs AV; Tse BW; Pearce AK; Yeh MC; Fletcher NL; Huang SS; Heston WD; Whittaker AK; Russell PJ; Thurecht KJ Evaluation of Polymeric Nanomedicines Targeted to Psma: Effect of Ligand on Targeting Efficiency. Biomacromolecules 2015, 16, 3235–3247. PubMed

Liu H; Rajasekaran AK; Moy P; Xia Y; Kim S; Navarro V; Rahmati R; Bander NH Constitutive and Antibody-Induced Internalization of Prostate-Specific Membrane Antigen. Cancer Res. 1998, 58, 4055–4060. PubMed

Tagawa ST; Milowsky MI; Morris M; Vallabhajosula S; Christos P; Akhtar NH; Osborne J; Goldsmith SJ; Larson S; Taskar NP; et al. Phase Ii Study of Lutetium-177-Labeled Anti-Prostate-Specific Membrane Antigen Monoclonal Antibody J591 for Metastatic Castration-Resistant Prostate Cancer. Clin. Cancer Res 2013, 19, 5182–5191. PubMed PMC

Tagawa ST; Milowsky MI; Morris MJ; Vallabhajosula S; Goldsmith S; Matulich D; Kaplan J; Berger F; Scher HI; Bander NH Phase Ii Trial of 177lutetium Radiolabeled Anti-Prostate-Specific Membrane Antigen (Psma) Monoclonal Antibody J591 (177lu-J591) in Patients (Pts) with Metastatic Castrate-Resistant Prostate Cancer (Metcrpc). J. Clin. Oncol 2008, 26, 5140. PubMed

Han DH; Wu JH; Han YH; Wei M; Han S; Lin RH; Sun ZY; Yang F; Jiao D; Xie P; et al. A Novel Anti-Psma Human Scfv Has the Potential to Be Used as a Diagnostic Tool in Prostate Cancer. Oncotarget 2016, 7, 59471–59481. PubMed PMC

Novakova Z; Foss CA; Copeland BT; Morath V; Baranova P; Havlinova B; Skerra A; Pomper MG; Barinka C Novel Monoclonal Antibodies Recognizing Human Prostate-Specific Membrane Antigen (Psma) as Research and Theranostic Tools. Prostate 2017, 77, 749–764. PubMed PMC

Ray Banerejee S; Kumar V; Lisok A; Plyku D; Novakova Z; Wharram B; Brummet M; Barinka C; Hobbs RF; Pomper MG Evaluation of (111)in-Dota-5d3, a Surrogate Spect Imaging Agent for Radioimmunotherapy of Prostate-Specific Membrane Antigen. J. Nucl. Med 2019, 60, 400. PubMed PMC

Beck A; Goetsch L; Dumontet C; Corvaia N Strategies and Challenges for the Next Generation of Antibody Drug Conjugates. Nat. Rev. Drug Discovery 2017, 16, 315–337. PubMed

Lyon R Drawing Lessons from the Clinical Development of Antibody-Drug Conjugates. Drug Discovery Today: Technol 2018, 30, 105–109. PubMed

Nicolaou KC; Rigol S Total Synthesis in Search of Potent Antibody-Drug Conjugate Payloads. From the Fundamentals to the Translational. Acc. Chem. Res 2019, 52, 127. PubMed

Diamantis N; Banerji U Antibody-Drug Conjugates-an Emerging Class of Cancer Treatment. Br. J. Cancer 2016, 114, 362–367. PubMed PMC

Bae YH; Park K Targeted Drug Delivery to Tumors: Myths, Reality and Possibility. J. Controlled Release 2011, 153, 198–205. PubMed PMC

Hapuarachchige S; Kato Y; Artemov D Bioorthogonal Two-Component Drug Delivery in Her2(+) Breast Cancer Mouse Models. Sci. Rep 2016, 6, 1. PubMed PMC

Hapuarachchige S; Zhu W; Kato Y; Artemov D Bioorthogonal, Two-Component Delivery Systems Based on Antibody and Drug-Loaded Nanocarriers for Enhanced Internalization of Nanotherapeutics. Biomaterials 2014, 35, 2346–2354. PubMed PMC

McKay CS; Finn MG Click Chemistry in Complex Mixtures: Bioorthogonal Bioconjugation. Chem. Biol 2014, 21, 1075–1101. PubMed PMC

Karver MR; Weissleder R; Hilderbrand SA Bioorthogonal Reaction Pairs Enable Simultaneous, Selective, Multi-Target Imaging. Angew. Chem., Int. Ed 2012, 51, 920–922. PubMed PMC

Chen Z; Krishnamachary B; Penet MF; Bhujwalla ZM Acid-Degradable Dextran as an Image Guided Sirna Carrier for Cox-2 Downregulation. Theranostics 2018, 8, 1–12. PubMed PMC

Liu J; Kopeckova P; Buhler P; Wolf P; Pan H; Bauer H; Elsasser-Beile U; Kopecek J Biorecognition and Subcellular Trafficking of Hpma Copolymer-Anti-Psma Antibody Conjugates by Prostate Cancer Cells. Mol. Pharmaceutics 2009, 6, 959–970. PubMed PMC

Vercauteren D; Vandenbroucke RE; Jones AT; Rejman J; Demeester J; De Smedt SC; Sanders NN; Braeckmans K The Use of Inhibitors to Study Endocytic Pathways of Gene Carriers: Optimization and Pitfalls. Mol. Ther 2010, 18, 561–569. PubMed PMC

Shang L; Nienhaus K; Nienhaus GU Engineered Nanoparticles Interacting with Cells: Size Matters. J. Nanobiotechnol 2014, 12, 5. PubMed PMC

Yameen B; Choi WI; Vilos C; Swami A; Shi JJ; Farokhzad OC Insight into Nanoparticle Cellular Uptake and Intracellular Targeting. J. Controlled Release 2014, 190, 485–499. PubMed PMC

Zhu W; Okollie B; Artemov D Controlled Internalization of Her-2/ Neu Receptors by Cross-Linking for Targeted Delivery. Cancer Biol. Ther 2007, 6, 1960–1966. PubMed

Kundranda MN; Niu J Albumin-Bound Paclitaxel in Solid Tumors: Clinical Development and Future Directions. Drug Des., Dev. Ther 2015, 9, 3767–3777. PubMed PMC

Rabbani G; Ahn SN Structure, Enzymatic Activities, Glycation and Therapeutic Potential of Human Serum Albumin: A Natural Cargo. Int. J. Biol. Macromol 2019, 123, 979–990. PubMed

Karimi M; Bahrami S; Ravari SB; Zangabad PS; Mirshekari H; Bozorgomid M; Shahreza S; Sori M; Hamblin MR Albumin Nanostructures as Advanced Drug Delivery Systems. Expert Opin. Drug Delivery 2016, 13, 1609–1623. PubMed PMC

Larsen MT; Kuhlmann M; Hvam ML; Howard KA Albumin-Based Drug Delivery: Harnessing Nature to Cure Disease. Mol. Cell Ther 2016, 4, 3. PubMed PMC

Umbricht CA; Benesova M; Schibli R; Muller C Preclinical Development of Novel Psma-Targeting Radioligands: Modulation of Albumin-Binding Properties to Improve Prostate Cancer Therapy. Mol. Pharmaceutics 2018, 15, 2297–2306. PubMed

Darko A; Wallace S; Dmitrenko O; Machovina MM; Mehl RA; Chin JW; Fox JM Conformationally Strained Trans-Cyclooctene with Improved Stability and Excellent Reactivity in Tetrazine Ligation. Chem. Sci 2014, 5, 3770–3776. PubMed PMC

Widdison WC; Wilhelm SD; Cavanagh EE; Whiteman KR; Leece BA; Kovtun Y; Goldmacher VS; Xie H; Steeves RM; Lutz RJ; et al. Semisynthetic Maytansine Analogues for the Targeted Treatment of Cancer. J. Med. Chem 2006, 49, 4392–4408. PubMed

Liebmann JE; Cook JA; Lipschultz C; Teague D; Fisher J; Mitchell JB Cytotoxic Studies of Paclitaxel (Taxol) in Human Tumour Cell Lines. Br. J. Cancer 1993, 68, 1104–1109. PubMed PMC

Development and therapeutic evaluation of 5D3(CC-MLN8237)3.2 antibody-theranostic conjugates for PSMA-positive prostate cancer therapy

Engineered Fragments of the PSMA-Specific 5D3 Antibody and Their Functional Characterization

Development of 5D3-DM1: A Novel Anti-Prostate-Specific Membrane Antigen Antibody-Drug Conjugate for PSMA-Positive Prostate Cancer Therapy