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

Abstract

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 89 Zr-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 89 Zr to obtain [ 89 Zr]ZrDFOamivantamab. Binding of the bispecific [ 89 Zr]ZrDFO-amivantamab as well as its mono-specific "single-arm" antibody controls were determined in vitro and in vivo. Biodistribution studies of [ 89 Zr]ZrDFO-amivantamab were performed in MDA-MB-468 xenografts to determine the optimal imaging time point. PET/CT imaging with [ 89 Zr]ZrDFO-amivantamab or its isotype control was performed in a panel of TNBC xenografts with varying levels of EGFR and c-MET expression. Results [ 89 Zr]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. [ 89 Zr]ZrDFO-amivantamab demonstrated bispecific binding in cell lines co-expressed with EGFR and c-MET. PET/CT imaging with [ 89 Zr]ZrDFO-amivantamab in TNBCxenografted 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, [ 89 Zr]ZrDFO-amivantamab, and evaluated its pharmacologic and imaging properties in comparison with its single-arm antibodies and non-specific isotype controls.[ 89 Zr]ZrDFO-amivantamab demonstrated the greatest uptake in tumors co-expressing EGFR and c-MET.

Preclinical Advances in Theranostics for the Different Molecular Subtypes of Breast Cancer

“…One of the reasons for their success is that bispecific constructs targeting two receptors can help to overcome drug resistance associated with mono-targeted therapies (Thakur et al, 2018). Different bispecific imaging and/or theranostic agents have also been developed, notably the scaffolds targeting GRPR/FA and EGFR/c-MET with 99m Tc/ 177 Lu-BBN-FA (Aranda-Lara et al, 2016a;Aranda-Lara et al, 2016b) and [ 89 Zr]ZrDFOamivantamab (Cavaliere et al, 2020), respectively. These agents hold potential for clinical translation due to the high expression of targets in several molecular subtypes of breast cancer and the promise to overcome resistance to monotargeted therapy due to their multiple mechanisms of action.…”

“…Amivantamab is a new bispecific antibody with multiple mechanisms of action, including inhibition of the EGFR and the hepatocyte growth factor receptor (HGFR/c-MET) pathways (Moores et al, 2016). In a recent report it was radiolabeled with 89 Zr via a desferrioxamine chelate (DFO) to create a companion diagnostic imaging agent for this bispecific antibody (Cavaliere et al, 2020) (Bispecific antibodies, Figure 2). As overexpression of EGFR and c-MET was found in TNBC and associated with progression of the disease, the resulting [ 89 Zr]ZrDFO-amivantamab radioconjugate was evaluated in TNBC xenograft models (Cavaliere et al, 2020;Chae et al, 2016).…”

“…In a recent report it was radiolabeled with 89 Zr via a desferrioxamine chelate (DFO) to create a companion diagnostic imaging agent for this bispecific antibody (Cavaliere et al, 2020) (Bispecific antibodies, Figure 2). As overexpression of EGFR and c-MET was found in TNBC and associated with progression of the disease, the resulting [ 89 Zr]ZrDFO-amivantamab radioconjugate was evaluated in TNBC xenograft models (Cavaliere et al, 2020;Chae et al, 2016). Three xenografts were used, MDA-MB-468, MDA-MB-231, and MDA-MB-453, which are characterized by high, moderate, and negative co-expression of EGFR and c-MET, respectively (Figure 5).…”

“…Three xenografts were used, MDA-MB-468, MDA-MB-231, and MDA-MB-453, which are characterized by high, moderate, and negative co-expression of EGFR and c-MET, respectively (Figure 5). PET/CT imaging with [ 89 Zr]ZrDFOamivantamab showed its ability to detect graded levels of EGFR and c-MET with standard uptake values (SUV mean ) of 6.0 ± 1.1, 4.2 ± 1.4, 1.5 ± 1.4 96 h p. i. in MDA-MB-468, MDA-MB-231, and MDA-MB-453, respectively (Figure 5) (Cavaliere et al, 2020). Further, tumor uptake of [ 89 Zr]ZrDFO-amivantamab was significantly higher than those of the radiolabeled single-arm parent antibodies [ 89 Zr] ZrDFO-α-EGFR or [ 89 Zr]ZrDFO-α-c-MET.…”

Radionuclide-Based Imaging of Breast Cancer: State of the Art

“…Additionally, amivantamab is a novel bispecific antibody that simultaneously targets EGFR and the hepatocyte growth factor receptor (HGFR/c-MET) that are overexpressed in TNBC. In a recent report, Cavaliere et al [ 213 ] radiolabeled amivantamab with 89 Zr, and it demonstrated higher tumor uptake than that of the radiolabeled single-arm parent antibodies.…”