Pharmacokinetics and Biodistribution of 89 Zr-Miltuximab and Its Antibody Fragments as Glypican-1 Targeting Immuno-PET Agents in Glioblastoma

• Published in: Molecular pharmaceutics Vol. 20; no. 3; pp. 1549 - 1563
• Main Authors: Ghosh, Saikat, Fletcher, Nicholas L, Huda, Pie, Houston, Zachary H, Howard, Christopher B, Lund, Maria E, Lu, Yanling, Campbell, Douglas H, Walsh, Bradley J, Thurecht, Kristofer J
• Format: Journal Article
• Language: English
• Published: United States 06.03.2023
• Subjects: Antibodies, Monoclonal - pharmacokinetics; Cell Line, Tumor; Glioblastoma; Glypicans; Humans; Immunoglobulin Fragments; Neoplasm Recurrence, Local; Positron-Emission Tomography - methods; Tissue Distribution; Zirconium; pharmacokinetics; antibody fragments; biodistribution; glioblastoma; glypican-1; PET
• ISSN: 1543-8384; 1543-8392
• DOI: 10.1021/acs.molpharmaceut.2c00760

Glioblastoma (GBM) is the most aggressive form of primary brain cancer, accounting for about 85% of all primary central nervous system (CNS) tumors. With standard treatment strategies like surgery, radiation, and chemotherapy, the median survival time of patients with GBM is only 12-15 months from diagnosis. The poor prognosis of GBM is due to a very high tumor recurrence rate following initial treatment, indicating a dire need for improved diagnostic and therapeutic alternatives for this disease. Antibody-based immunotheranostics holds great promise in treating GBM, combining the theranostic applications of radioisotopes and target-specificity of antibodies. In this study, we developed and validated antibody-based positron emission tomography (PET) tracers targeting the heparan sulfate proteoglycan, glypican-1 (GPC-1), for noninvasive detection of disease using diagnostic molecular imaging. GPC-1 is overexpressed in multiple solid tumor types, including GBM, and is a promising biomarker for novel immunotheranostics. Here, we investigate zirconium-89 ( Zr)-conjugated Miltuximab (a clinical stage anti-GPC-1 monoclonal antibody developed by GlyTherix, Ltd.) and engineered fragments for their potential as immuno-PET tracers to detect GPC-1 GBM tumors in preclinical models. We explore the effects of molecular size, avidity, and Fc-domain on the pharmacokinetics and biodistribution in vivo, by comparing in parallel the full-length antibody (Miltuximab), Fab'2, Fab, and single-chain variable fragment (scFv) formats. High radiolabeling efficiency (>95%) was demonstrated by all the formats and the stability post-radiolabeling was higher for larger constructs of Miltuximab and the Fab. Receptor-mediated internalization of all Zr-labeled formats was observed in a human GBM cell line in vitro, while full-length Miltuximab demonstrated the highest tumor retention (5.7 ± 0.94% ID/g, day-9 postinjection (p.i.)) and overall better tumor-to-background ratios than the smaller Fc-less formats. Results from in vivo PET image quantification and ex vivo scintillation counting were highly correlated. Altogether, Zr-DFO-Miltuximab appears to be an effective immuno-PET imaging agent for detecting GPC-1 tumors such as GBM and the current results support utility of the Fc containing whole mAb format over smaller antibody fragments for this target.