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Translational Therapeutics

Biparatopic anti-HER2 drug radioconjugates as breast cancer theranostics

British Journal of Cancer volume 129, pages 153–162 (2023)Cite this article

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Abstract

Background

HER2 is overexpressed in 25–30% of breast cancer. Multiple domains targeting of a receptor can have synergistic/additive therapeutic effects.

Methods

Two domain-specific ADCs trastuzumab-PEG6-DM1 (domain IV) and pertuzumab-PEG6-DM1 (domain II) were developed, characterised and radiolabeled to obtain [89Zr]Zr-trastuzumab-PEG6-DM1 and [67Cu]Cu-pertuzumab-PEG6-DM1 to study their in vitro (binding assay, internalisation and cytotoxicity) and in vivo (pharmacokinetics, biodistribution and immunoPET/SPECT imaging) characteristics.

Results

The ADCs had an average drug-to-antibody ratio of 3. Trastuzumab did not compete with [67Cu]Cu-pertuzumab-PEG6-DM1 for binding to HER2. The highest antibody internalisation was observed with the combination of ADCs in BT-474 cells compared with single antibodies or ADCs. The combination of the two ADCs had the lowest IC50 compared with treatment using the single ADCs or controls. Pharmacokinetics showed biphasic half-lives with fast distribution and slow elimination, and an AUC that was five-fold higher for [89Zr]Zr-trastuzumab-PEG6-DM1 compared with [67Cu]Cu-pertuzumab-PEG6-DM1. Tumour uptake of [89Zr]Zr-trastuzumab-PEG6-DM1 was 51.3 ± 17.3% IA/g (BT-474), and 12.9 ± 2.1% IA/g (JIMT-1) which was similarly to [67Cu]Cu-pertuzumab-PEG6-DM1. Mice pre-blocked with pertuzumab had [89Zr]Zr-trastuzumab-PEG6-DM1 tumour uptakes of 66.3 ± 33.9% IA/g (BT-474) and 25.3 ± 4.9% IA/g (JIMT-1) at 120 h p.i.

Conclusion

Using these biologics simultaneously as biparatopic theranostic agents has additive benefits.

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Fig. 1: Flow cytometry saturation binding assay of antibodies and immunoconjugates in HER2-positive BT-474 cells.
Fig. 2: Radioligand and competitive binding assays in HER2-positive BT-474 cells.
Fig. 3: Antibody internalisation of trastuzumab, trastuzumab-PEG6-DM1, pertuzumab, pertuzumab-PEG6-DM1 and a combination of trastuzumab-PEG6-DM1 + pertuzumab-PEG6-DM1 in HER2-positive BT-474 cell line after 48 h of incubation.
Fig. 4: In vitro cytotoxicity of immunoconjugates in cells with different HER2 densities.
Fig. 5: MicroPET/SPECT/CT and biodistribution of antibody–drug radioconjugates at different timepoints post injection.
Fig. 6: Targeted antibody drug radioconjugate-specific imaging of HER2-positive xenografts following simultaneous injection of [89Zr]Zr-trastuzumab-PEG6-DM1 + [67Cu]Cu-pertuzumab-PEG6-DM1.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors wish to acknowledge the contributions of Dr. Vijay Gaja (Canadian Isotope Innovations Corp CIIC) for providing [67Cu]CuCl2, and Dr. Musharraf Khan (Saskatchewan Cyclotron Facility) for assistance with experiments.

Funding

This work was funded by Canadian Institute for Health Research (CIHR) Project Grants (# 437660 and 408132) to HF.

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Authors and Affiliations

  1. Department of Medical Imaging, College of Medicine, University of Saskatchewan, Saskatoon, SK, S7N 0W8, Canada

    Jessica Pougoue Ketchemen, Hanan Babeker, Anjong Florence Tikum, Anand Krishnan Nambisan, Fabrice Ngoh Njotu, Emmanuel Nwangele & Humphrey Fonge

  2. Department of Pathology and Lab. Medicine, College of Medicine, University of Saskatchewan, 107 Wiggins Rd, Saskatoon, SK, S7N 5A2, Canada

    Hanan Babeker

  3. Department of Medical Imaging, Royal University Hospital Saskatoon, SK, Saskatoon, SK, S7N 0W8, Canada

    Humphrey Fonge

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Contributions

Experimental design, execution and data analysis were performed by JPK, HB, AFT, AKN, FNN, EN and HF. Writing of the original draft preparation and review were done by JPK and HF. All the authors contributed to the article and approved the submitted version.

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Correspondence to Humphrey Fonge.

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The authors declare no competing interests.

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All animal experiments were approved, supervised, and maintained following the guidelines of the University of Saskatchewan Animal Care Committee (UACC). Ethical approval references 20170084 and 20220021.

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Ketchemen, J.P., Babeker, H., Tikum, A.F. et al. Biparatopic anti-HER2 drug radioconjugates as breast cancer theranostics. Br J Cancer 129, 153–162 (2023). https://doi.org/10.1038/s41416-023-02272-4

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