The goal of this competitive renewal application is to increase the clinical utility of labeled mAbs for cancer diagnosis and therapy by developing more effective strategies for labeling internalizing mAbs and mAb fragments with radioiodine nuclides and 211At. With the emergence of the epidermal growth factor receptor variant III (EGFRvIII) as a tumor-specific molecular target on glioma and other tumors, research during the previous grant period was focused on the development of strategies for labeling mAbs reactive with this rapidly internalizing mutant receptor. Our studies revealed that radio iodination of anti-EGFRvIII mAbs using reagents containing benzoates bearing charged substituent's or D-amino acid peptides significantly improved retention of radioactivity in EGFRvIII-expressing tumor cells and xenografts compared with mAbs labeled by conventional approaches. The most promising results were obtained with N5-(3-[*I]iodobenzoyl)-Lys5-N1-maleimido-D-Gly1- GEEEK ([*I]IB-Mal-D-GEEEK), which includes 3 D-glutamates to provide a negatively charged, proteolytically inert moiety and an iodobenzoyl group to minimize dehalogenation. Our hypothesis is that optimized labeling methods for internalizing mAbs such as anti-EGFRvIII based on the IB-Mal-D-GEEEK template will enhance tumor retention and tumor-to-normal tissue ratios, thereby improving their clinical potential as diagnostic and therapeutic agents. We propose to: 1) label anti-EGFRvIII mAbs and fragments with radioiodine nuclides and 211At using IB-Mal-D-GEEEK and its [211At]astatobenzoyl Mal-D-GEEEK analogue and to evaluate their potential as diagnostic and therapeutic radiopharmaceuticals;2) investigate strategies for improving the Mal- GEEEK reagent for labeling internalizing mAbs including alteration in D-peptide sequence (number and nature of negatively charged amino acids), use of a less hydrophobic dehalogenation resistant prosthetic group such as iodopyridine, incorporation of a cathepsin B cleavable linker, and coupling the radiohalogenation precursor to the mAb prior to the labeling reaction;3) investigate the nature of the low and high molecular weight labeled catabolites generated in tumor cells in vitro and tumor and normal tissues in vivo;4) evaluate the therapeutic efficacy of promising 131I- and 211At-labeled anti-EGFRvIII conjugates in athymic rodents with subcutaneous, intracranial, and neoplastic meningitis xenografts;and 5) With the best radio labeled anti-EGFRvIII conjugates, conduct all the toxicity, efficacy, dosimetry, and other FDA-required studies for Investigational New Drug permits, to all performance of clinical trials in malignant glioma patients under our Brain Tumor Center grant (NS20023

Public Health Relevance

Our goal is to develop methods for attaching iodine radio nuclides and 1-particle emitting 211At to monoclonal antibodies (mAbs) in such a way that the radioactivity remains trapped in the cancer cell after cellular metabolism of mAb. Although the proposed research is focused on improving the imaging and treatment of brain tumors and mAbs that target EGFRvIII, these labeled mAbs also might be useful in the management of other cancers that over express this tumor-specific receptor. Furthermore, these labeling methods should be applicable to other internalizing mAbs and fragments, increasing the potential impact of this work.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01CA042324-28
Application #
8657792
Study Section
Clinical Molecular Imaging and Probe Development (CMIP)
Program Officer
Prasanna, Pat G
Project Start
Project End
Budget Start
Budget End
Support Year
28
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Duke University
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
City
Durham
State
NC
Country
United States
Zip Code
27705
Pruszynski, Marek; Koumarianou, Eftychia; Vaidyanathan, Ganesan et al. (2015) D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination. Nucl Med Biol 42:19-27
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Koumarianou, Eftychia; Slastnikova, Tatiana A; Pruszynski, Marek et al. (2014) Radiolabeling and in vitro evaluation of (67)Ga-NOTA-modular nanotransporter--a potential Auger electron emitting EGFR-targeted radiotherapeutic. Nucl Med Biol 41:441-9
Pruszynski, Marek; Koumarianou, Eftychia; Vaidyanathan, Ganesan et al. (2014) Improved tumor targeting of anti-HER2 nanobody through N-succinimidyl 4-guanidinomethyl-3-iodobenzoate radiolabeling. J Nucl Med 55:650-6
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Choi, Jaeyeon; Vaidyanathan, Ganesan; Koumarianou, Eftychia et al. (2014) N-Succinimidyl guanidinomethyl iodobenzoate protein radiohalogenation agents: influence of isomeric substitution on radiolabeling and target cell residualization. Nucl Med Biol 41:802-12
Pruszynski, Marek; Koumarianou, Eftychia; Vaidyanathan, Ganesan et al. (2013) Targeting breast carcinoma with radioiodinated anti-HER2 Nanobody. Nucl Med Biol 40:52-9
Zalutsky, Michael R; Boskovitz, Abraham; Kuan, Chien-Tsun et al. (2012) Radioimmunotargeting of malignant glioma by monoclonal antibody D2C7 reactive against both wild-type and variant III mutant epidermal growth factor receptors. Nucl Med Biol 39:23-34
Chitneni, Satish K; Palmer, Gregory M; Zalutsky, Michael R et al. (2011) Molecular imaging of hypoxia. J Nucl Med 52:165-8
Zalutsky, Michael R; Pruszynski, Marek (2011) Astatine-211: production and availability. Curr Radiopharm 4:177-85

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