The development of a highly specific radiochemical compound based on labeled 99mtc recombinant molecules for targeted imaging of cells with the overexpression of Her-2 / neu

Currently, there is a urgent need to search for new diagnostic methods that allow us to reveal malignant tumors with the overexpression of Her-2/neu with high accuracy. In recent years radioisotope methods have been actively developing to identify specific tumor targets, with antibodies being the “targeting” module.
The purpose of the study. Creation of a chemically stable radiochemical compound for the imaging of cells with the overexpression of Her-2/neu.
Materials and methods. The study was conducted using two human adenocarcinoma cell lines with expression (BT-474) and without expression (MCF-7) Her-2/neu. The specificity of the binding of the test complex with Her-2/neu receptor was determined by direct radiometric and planar scintigraphy. To evaluate the differences in quantitative characteristics between the groups a non-parametric Mann – Whitney test was used.
Results. The yield of the labeled complex was more than 91% and the radiochemical frequency was more than 94%. When performing a visual scintigraphic evaluation, a much higher accumulation rate of the studied radiopharmaceutical preparation (RFP) was observed in the culture of cells with overexpression of the surface Her-2/neu receptor. Direct radiometric results also demonstrated a higher accumulation of RFPs in the human BT-474 mammary adenocarcinoma cell line with Her-2/neu overexpression in comparison with the control group.
Conclusion. Preclinical studies demonstrated high stability of the test compound, as well as its accumulation in the group of cells with Her-2/neu overexpression

1. Polanovski O. L., Lebedenko E. N., Deyev S. M. ERBB oncogeni – misheni monoclonalnikh antitel [ERBB – oncogenes as targets for monoclonal antibodies] // Biokhimiya – Biochemistry. 2012; 3 (77): 289–311 (in Russian).

2. Chernov V.I., Bragina O.D., Sinilkin I.G., et al. Radioimmunoterapiya: sovremennoe sostoyanie problemi [Radioimmunotherapy: current state of the problem] // Voprosi Оncologii – Questions of Оncology. 2016; 62 (1): 24–30 (in Russian).

3. Chernov V.I., Bragina O.D., Sinilkin I.G. et al. Radioimmunotherapiya v lechenii zlokachestvennix obrazovanii [Radioimmunotherapy in the treatment of malignancies] // Sibirskii Oncologicheskii Jurnal – Siber. J. Oncol. 2016; 15 (2): 101–106 (in Russian).

4. Slamon D.J., Clark G.M., Wong S.G. et al. Human breast cancer: correlation of relapse an survival with amplification of the Her-2/neu oncogenes // Science. 1987; 235: 177–182. doi: 10.1126/science.3798106.

5. Romond E.H., Perez E.A., Bryant J. et al. Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer // N. Engl. J. Med. 2005; 353: 1673–1684. doi: 10.1056/NEJMoa052122.

6. Verma S., Miles D., Gianni L. et al. Trastuzumab emtansine for HER2-positive advanced breast cancer // N. Engl. J .Med. 2012; 367: 1783–1791. doi: 10.1056/NEJMoa1209124.

7. Babyshkina N., Malinovskaya E., Cherdinceva N. et al. Neoadjuvant chemotherapy for different molecular breast cancer subtypes: a retrospective study in Russian population // Medical Oncology. 2014; 31 (9): 1–12.

8. Chernov V.I., Bragina O.D., Sinilkin I.G. et al. Radionuclidnaya teranostica zlokachestvennix obrazovanii [Radionuclide teranostic of malignancies] // Vestnik Rentgenologii i Radiologii – Vestnik Rentgenologii and Radiologii. 2016; 97 (5): 306–313 (in Russian).

9. Zahid M., Khan S., Khan R. et al. Detection of HER2/ neu gene amplification by fluoroscence in situ hybridization technique // Pathology. 2016; 48 (1): 163–170.

10. Orlando L., Viale G., Bria E. et al. Discordance in pathology report after central pathology review: Implications for breast cancer adjuvant treatment // Breast. 2016; 30: 151–155. doi: 10.1016/j.breast.2016.09.015.

11. Telugu R.B., Chowhan A.K., Rukmangadha N. et al. Human epidermal growth factor receptor 2/neu protein expression in meningiomas: An immunohistochemical study // J. Neurosci Rural Pract. 2016; 7 (4): 526–531. doi: 10.4103/0976-3147.188640.

12. Hanna W.M., Rüschoff J., Bilous M. et al. HER2 in situ hybridization in breast cancer: clinical implications of polysomy 17 and genetic heterogeneity // Mod. Pathol. 2013; 27: 4–18. doi: 10.1038/modpathol.2013.103.

13. Kurozumi1 S., Padilla M., Kurosumi M. et al. HER2 intratumoral heterogeneity analyses by concurrent HER2 gene and protein assessment for the prognosis of HER2 negative invasive breast cancer patients // Breast Cancer Res Treat. 2016; 158: 99–111. doi: 10.1007/s10549-016-3856-2.

14. Seol H., Lee H.J., Choi Y. et al. Intratumoral heterogeneity of HER2 gene amplification in breast cancer: its clinicopathological significance // Modern Pathology. 2012; 25: 938–948. doi: 10.1038/modpathol.2012.36.

15. Riccarbona G., Decristoforo C. Peptide targeted imaging of cancer // Cancer Biother. Radiopharm. 2003; 18: 675–687. doi:10.1089/108497803770418238.

16. Engfeldt T., Orlova A., Tran T. et al. Imaging of HER2-expressing tumours using a synthetic Affibody molecule containing the 99mTc-chelating mercaptoacetyl-glycyl-glycyl-glycyl (MAG3) sequence // European Journal of Nuclear Medicine and Molecular Imaging. 2007; 34 (5). 722–733. doi: 10.1007/s00259-006-0266-4.

17. Stumpp M. T., Binz H.K., Amstutz P. DARPins: A new generation of protein therapeutics // Drug Discovery Today. 2008; 13 (15): 695–701.

18. Tamaskovic R., Simon M., Stefan N. et al. Designed ankyrin repeat proteins (DARPins) from research to therapy // Methods Enzymol. 2012; 503: 101–134. doi: 10.1016/j.drudis.2008.04.013.

19. Boersma Y.L., Pluckthun A. DARPins and other repeat protein scaffolds: advances in engineering and applications // Curr. Opin. Biotechnol. 2011; 22: 849–57. doi: 10.1016/j.copbio.2011.06.004.

20. Binz H.K., Stumpp M.T., Forrer P., Amstutz P., Pluckthun A. Designing repeat proteins: well-expressed, soluble and stable proteins from combinatorial libraries of consensus ankyrin repeat proteins // J. Mol. Biol. 2003; 332: 489–503.

21. Skuridin V.S., Stasyuk E.S., Nesterov E.A., Sadkin V.L., Rogov A.S. Adsorption of 99mTc on Aluminum Oxide // Radiochemistry. 2011; 5 (53): 448–451.

22. Stasyuk E.S., Nesterov E.A., Skuridin V.S. et al. A procedure for sorbent pretreatment for the production of high-activity 99Mo/99mTc generators based on enriched 98Mo // Radiochemistry. 2012; 54 (4): 391–394.

23. Gabriel M., Decristoforo C., Donnemiller E. et al. An intrapatient comparison of 99mTc-EDDA/HYNICTOC with 111In-DTPA-octreotide for diagnosis of somatostatin receptorexpressing tumors // J. Nucl. Med. 2003; 44: 708–716.

24. Yusubov M.S., Svitich D.Y., Larkina M.S. et al. Applications of iodonium salts and iodonium ylides as precursors for nucleophilic fluorination in Positron Emission Tomography // ARKIVOC. 2013; (i): 364–395.

25. Kulibaba E.V., Larkina M.S. New syntheses of bis(2-pyridylmethyl)amino acid // IX international conference of young scientists on chemistry «Mendeleev 2015». Book of abstracts. Saint Petersburg. 7–10 of April 2015; 250.

26. Skuridin V.S., Stasyuk E.S., Rogov A.S. et al. Modified DTPA molecule-based nanocolloid radiopharmaceuticals // Journal of Radioanalytical and Nuclear Chemistry. 2015; 303 (3): 1961–1965.

27. Yusubov M.S., Jdankin V.V., Larkina M.S. et al. Sposob polucheniya ω- iodalifalicheskix karbonovix kislot I ikh efirov [Method of producing omega-iodo-aliphatic carboxylic acids and esters thereof]. Patent № 2494087. 2013; 27.09.2013 (in Russian).