Development of [89Zr]ZrDFO-amivantamab bispecific to EGFR and c-MET for PET imaging of triple-negative breast cancer

Background: Amivantamab is a novel bispecific antibody that simultaneously targets the epidermal growth factor receptor (EGFR) and the hepatocyte growth factor receptor (HGFR/c-MET) that are overexpressed in several types of Cancer including triple-negative breast Cancer (TNBC). Targeting both receptors simultaneously can overcome resistance to mono-targeted therapy. The purpose of this study is to develop ⁸⁹Zr-labeled amivantamab as a potential companion diagnostic imaging agent to amivantamab therapy using various preclinical models of TNBC for evaluation.

Methods: Amivantamab was conjugated to desferrioxamine (DFO) and radiolabeled with ⁸⁹Zr to obtain [⁸⁹Zr]ZrDFO-amivantamab. Binding of the bispecific [⁸⁹Zr]ZrDFO-amivantamab as well as its mono-specific "single-arm" antibody controls were determined in vitro and in vivo. Biodistribution studies of [⁸⁹Zr]ZrDFO-amivantamab were performed in MDA-MB-468 xenografts to determine the optimal imaging time point. PET/CT imaging with [⁸⁹Zr]ZrDFO-amivantamab or its isotype control was performed in a panel of TNBC xenografts with varying levels of EGFR and c-MET expression.

Results: [⁸⁹Zr]ZrDFO-amivantamab was synthesized with a specific activity of 148 MBq/mg and radiochemical yield of ≥ 95%. Radioligand binding studies and western blot confirmed the order of EGFR and c-MET expression levels: HCC827 lung Cancer cell (positive control) > MDA-MB-468 > MDA-MB-231 > MDA-MB-453. [⁸⁹Zr]ZrDFO-amivantamab demonstrated bispecific binding in cell lines co-expressed with EGFR and c-MET. PET/CT imaging with [⁸⁹Zr]ZrDFO-amivantamab in TNBC xenografted mice showed standard uptake value (SUV_mean) of 6.0 ± 1.1 in MDA-MB-468, 4.2 ± 1.4 in MDA-MB-231, and 1.5 ± 1.4 in MDA-MB-453 tumors, which are consistent with their receptors' expression levels on the cell surface.

Conclusion: We have successfully prepared a radiolabeled bispecific antibody, [⁸⁹Zr]ZrDFO-amivantamab, and evaluated its pharmacologic and imaging properties in comparison with its single-arm Antibodies and non-specific isotype controls. [⁸⁹Zr]ZrDFO-amivantamab demonstrated the greatest uptake in tumors co-expressing EGFR and c-MET.

89Zr; Bispecific antibody; C-MET; EGFR; Triple-negative breast cancer.