Development and evaluation of 89Zr-trastuzumab for clinical applications

Document Type : Original Article

Authors

1 Radiation Application Research School, Nuclear Science and Technology Research Institute, Tehran, Iran

2 Khatam PET/CT Center, Khatam-Al-Anbia Hospital, Tehran, Iran

3 Department of Clinical Biochemistry, Faculty of Medical Sciences, Zanjan University Medical Sciences, Zanjan, Iran

Abstract

Objective(s): Due to the suitable physical characteristics of 89Zr as a PET radionuclide and affinity of Trastuzumab monoclonal antibody against HER2, [89Zr]Zr-Trastuzumab was prepared and went through preclinical evaluations for ultimate human applications.
Methods: 89Zr was produced by using 89Y(p,n)89Zr reaction at a 30 MeV cyclotron (radionuclide purity>99.9%, specific activity of 17 GBq/µg). p-SCN-Bn-Deferoxamine (DFO); was conjugated to trastuzumab, followed by labeling with 89Zr in oxalate form at optimized condition. Cell binding, internalization and, radioimmuno-activity assays were studied using HER2+ BT474 and HER2- CHO cell lines. Finally, the biodistribution of the radioimmunoconjugate was assessed in normal and HER2+ BT474 tumor-bearing mice using tissue counting and imaging at different intervals after injection. Also, a woman with HER2-positive metastatic breast cancer under treatment with Herceptin underwent both [89Zr]Zr-Trastuzumab and, [18F]FDG PET/CTs.
Results: 89Zr was produced with high radionuclidic and radiochemical purities (>99%) and [89Zr]Zr-DFO-Trastuzumab was prepared with radiochemical purity of >98% and specific activity of 9.85 GBq/µmol. The radioimmunoconjugate was stable both in PBS buffer and in human serum for at least 48 h. The radioimmunoactivity assay demonstrated about 70% of [89Zr]Zr-DFO-Trastuzumab is bound to the BT474 cells at the number of 250×106 cells. Cell binding studies showed that about 28% of radioimmunoconjugate is attached to BT474 cells after 90 min. Internalization studies showed that 50% of [89Zr]Zr-Trastuzumab is internalized to BT474 cells only in 6 h. The biodistribution study of the labeled compound in normal mice demonstrated the same pattern of the monoclonal antibodies which is entirely different from the biodistribution of free 89Zr. Biodistribution and imaging studies in tumor-bearing mice showed the significant uptake values of [89Zr]Zr-Trastuzumab in tumor sites. [89Zr]Zr-Trastuzumab PET/CT revealed metastatic lesions documented previously with [18F]FDG PET/CT scan in a woman with breast cancer who was under treatment with Herceptin. Although the [18F]FDG PET/CT scan had better quality images, the valuable and unique advantage of [89Zr]Zr-Trastuzumab PET/CT is delineating HER2+ metastasis, which is essential in diagnosis and HER2-based treatments.
Conclusion: The prepared [89Zr]Zr-Trastuzumab has a high potential radio-pharmaceutical for immune-PET imaging of the patients with HER2+ tumors.

Keywords

Main Subjects

References

  1. Iqbal N, Iqbal N. Human Epidermal Growth Factor Receptor 2 (HER2) in Cancers: Overexpression and Therapeutic Implications. Mol Biol Int. 2014; 852748.
  2. Massicano AVF, Lee S, Crenshaw BK, Aweda TA, El Sayed R, Super I, et al. Imaging of HER2 with [89Zr]pertuzumab in Response to T-DM1 Therapy. Cancer Biother Radiopharm. 2019; 34(4):209-217.
  3. McKeage K, Perry CM. Trastuzumab: a review of its use in the treatment of metastatic breast cancer overexpressing HER2. Drugs. 2002; 62(1):209-43.
  4. Signore A, Lauri C, Auletta S, Varani M, Onofrio L, Glaudemans AWJM, et al. Radiopharmaceuticals for Breast Cancer and Neuroendocrine Tumors: Two Examples of How Tissue Characterization May Influence the Choice of Therapy. Cancers (Basel). 2020; 12(4):781.
  5. Abbas Abadi S, Alirezapour B, Kertész I, Rasaee MJ, Mohammadnejad J, Paknejad M, et al. Preparation, quality control, and biodistribution assessment of [111In]In-DOTA-PR81 in BALB/c mice bearing breast tumors. J Labelled Comp Radiopharm. 2021; 64(4):168-180.
  6. Zolghadri S, Naderi M, Yousefnia H, Alirezapour B, Beiki D. Evaluation of the Possible Utilization of 68Ga-DOTATOC in Diagnosis of Adenocarcinoma Breast Cancer. Asia Ocean J Nucl Med Biol. 2018; 6(1):41-49.
  7. Wang RE, Zhang Y, Tian L, Cai W, Cai J. Antibody-based imaging of HER-2: moving into the clinic. Curr Mol Med. 2013; 13(10): 1523-37.
  8. Alirezapour B , Jalilian AR , Rajabifar S , Mirzaii M , Moradkhani S , Pouladi M , et al. Preclinical evaluation of [111In]-DOTA-trastuzumab for clinical trials. J Cancer Res Ther. 2014; 10(1): 112-20.
  9. Beylergil V, Morris PG, Smith-Jones PM, Modi S, Solit D, Hudis CA, et al. Pilot study of 68Ga-DOTA-F(ab')2-trastuzumab in patients with breast cancer. Nucl Med Commun. 2013; 34(12):1157-65.
  10. Pandey MK, Engelbrecht HP, Byrne JP, Packard AB, DeGrado TR. Production of 89Zr via the 89Y(p,n)89Zr reaction in aqueous solution: Effect of solution composition on in-target chemistry. Nucl Med Biol. 2014; 41(4): 309–16.
  11. Ciarmatori A, Cicoria G, Pancaldi D, Infantino A, Boschi S, Fanti S, et al. Some experimental studies on 89Zr production. Radiochim. Acta. 2011; 99: 631–4.
  12. Jauw YW, Menke-van der Houven van Oordt CW, Hoekstra OS, Hendrikse NH, Vugts DJ, Zijlstra JM, et al. Immuno-Positron Emission Tomography with Zirconium-89-Labeled Monoclonal Antibodies in Oncology: What Can We Learn from Initial Clinical Trials? Front Pharmacol. 2016; 7: 131.
  13. Dijkers EC, Kosterink JG, Rademaker AP, Perk LR, van Dongen GA, Bart J, et al. Development and characterization of clinical-grade 89Zr-trastuzumab for HER2/ neu immunoPET imaging. J Nucl Med. 2009; 50:974–81.
  14. Dijkers EC, Oude Munnink TH, Kosterink JG, Brouwers AH, Jager PL, de Jong JR, et al. Bio-distribution of 89Zr-trastuzumab and PET imaging of HER2- positive lesions in patients with metastatic breast cancer. Clin Pharmacol Ther. 2010; 87(5):586-92.
  15. Laforest R, Lapi SE, Oyama R, Bose R, Tabchy A, Marquez-Nostra BV, et al. [89Zr]Trastuzumab: Evaluation of Radiation Dosimetry, Safety, and Optimal Imaging Parameters in Women with HER2-Positive Breast Cancer. Mol Imaging Biol. 2016; 18(6):952-9.
  16. Vosjan MJ, Perk LR, Visser GW, Budde M, Jurek P, Kiefer GE, et al. Conjugation and radiolabeling of monoclonal antibodies with zirconium-89 for PET imaging using the bifunctional chelate p-isothiocyanatobenzyl- Nat Protoc. 2010; 5(4): 739-43.
  17. Lindmo T, Boven E, Cuttitta F, Fedorko J, Bunn PA Jr. Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J Immunol Methods. 1984; 72(1): 77-89.
  18. Marquez BV, Ikotun OF, Zheleznyak A, Wright B, Hari-Raj A, Pierce RA, et al. Evaluation of (89)Zr-pertuzumab in Breast cancer xenografts. Mol Pharm. 2014; 11(11):3988-95.
  19. Chang AJ, Desilva R, Jain S, Lears K, Rogers B, Lapi S. 89Zr-Radiolabeled Trastuzumab Imaging in Orthotopic and Metastatic Breast Tumors. Pharmaceuticals (Basel). 2012; 5(1):79-93.
  20. Raavé R, Sandker G, Adumeau P, Jacobsen CB, Mangin F, Meyer M, et al. Direct comparison of the in vitro and in vivo stability of DFO, DFO* and DFOcyclo* for 89Zr-immunoPET. Eur J Nucl Med Mol Imaging. 2019; 46(9):1966-77.
  21. Yousefnia H, Radfar E, Jalilian AR, Bahrami-Samani A , Shirvani-Arani S, Arbabi A, et al. Development of 177Lu-DOTA-anti-CD20 for radioimmunotherapy. J Radioanal Nucl Chem. 2011; 287 (1): 199-209.
  22. Bahrami-Samani A, Ghannadi-Maragheh M, Jalilian AR, Yousefnia H, Garousi J, Moradkhani M. Development of 153Sm-DTPA-rituximab for radioimmunotherapy. Nukleonika. 2009; 54 (4): 271-7.
Asia Oceania Journal of Nuclear Medicine and Biology
Volume 11, Issue 2
June 2023
Pages 135-144

Files

Share

How to cite

Statistics