Synthesis of 111In-p-SCN-Bn-DTPA-nimotuzumab and its preclinical evaluation in EGFR positive NSCLC animal model

Gul-e Raana; Syed Qaiser Shah

doi:10.1515/ract-2021-1054

Article

Synthesis of ¹¹¹In-p-SCN-Bn-DTPA-nimotuzumab and its preclinical evaluation in EGFR positive NSCLC animal model

Gul-e Raana and Syed Qaiser Shah

Published/Copyright: October 20, 2021

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From the journal Radiochimica Acta Volume 109 Issue 12

Abstract

The aim of this study was to investigate labeling of nimotuzumab (h-R3) with ¹¹¹In using p-SCN-Bn-DTPA as bifunctional chelate, evaluate its targeting potential against SK-LU-1, H226, H650, H661, and HCC4006 non-small cell lung carcinoma (NSCLC) cell lines and correlate epidermal growth factor receptor (EGFR) expression level with internalization kinetics, biodistribution and imaging accuracy using Balb/c mice and New Zealand White rabbit (NZWR) animal model. The amount of p-SCN-Bn-DTPA attached to h-R3 was assessed by measuring relative absorbance at 652 nm with ultraviolet (UV) spectrophotometer. High-performance liquid chromatography (HPLC) was used to determine percent radiochemical purity (%RCP) and in vitro stability using excess amount of diethylenetriamine pentaacetate (DTPA). The in vitro stability in rat serum was estimated using iTLC-SG. EGFR expression level in each tumor was assessed by chemiluminescence. In vivo uptake in different organs of Balb/c mice and non-invasive imaging potential using NZWR bearing HCC4006 tumor, was evaluated with gamma camera. UV spectroscopy has confirmed the attachment of five p-SCN-Bn-DTPA (chelate) with one antibody. The HPLC indicated 98.85 ± 0.14% (n = 3) %RCP with high yield (>96%), specific activity 3.5 ± 0.0.25 mCi per mg and 94.25 ± 0.34% in vitro stability at 37 °C in mice serum. In excess DTPA no considerable transchelation was experiential from the ¹¹¹In labeled p-SCN-Bn-DTPA-h-R3 to the challenger. The EGFR expression in HCC4006 was higher amongst all with band density of 23.53 relative to 1.00 of H226. Initially internalization was lower which went up 1.05 × 10⁴ molecules per HCC4006 cell in 48 h. The optimal concentration of h-R3 for maximum uptake was 15 μg per animal. Higher uptake in target organ was observed in animal infected with HCC4006 cells. However, in excess pure h-R3 the uptake was significantly reduced indicating tumor specificity. HCC4006 target site was undistinguishable relative to background activity in the initial phase of imaging due to poor uptake. However, within 60 h the HCC4006 tumor was quite apparent. This experiment suggests that at optimal dosage of ¹¹¹In labeled h-R3 can be used for localization and identification of EGFR positive NSCLC using gamma camera.

Keywords: 111In labeled h-R; biodistribution ad imaging; EGFR; non-small cell lung carcinoma (NSCLC)

Corresponding author: Gul-e-Raana, Biochemistry & Nuclear Medicine Research Laboratory, Institute of Chemical Sciences, University of Peshawar, Peshawar, 25120 K.P.K, Pakistan, E-mail: guleraanachem04@gmail.com

Funding source: Higher Education Commission (HEC)

Award Identifier / Grant number: 3122

Acknowledgments

We thank the Higher Education Commission (HEC), Islamabad, Pakistan for providing all sorts of financial assistance under National Research Program for Universities. The instant study is part of the HEC funded research grant No. 3122.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: The study was supported by Higher Education Commission (HEC), Islamabad, Pakistan under National Research Program for Universities (grant no. 3122).
Conflict of interest statement: The authors declare no conflicts of interest regarding this article.
Ethics approval and consent to participate: No human was involved in this study. Experiments on mice and rabbits models were executed in compliance with Nuclear Medicine Research Laboratory (NMRL) and approval of ethics committee at Institute of Chemical Sciences (ICS), University of Peshawar.
Consent for publication: Not applicable.
Availability of data and materials: Not applicable.

References

1. Duma, N., Santana-Davila, R., Molina, J. R. Non-small cell lung cancer: epidemiology, screening, diagnosis, and treatment. Mayo Clin. Proc. 2019, 94, 1623; https://doi.org/10.1016/j.mayocp.2019.01.013.Search in Google Scholar PubMed

2. Vaidya, P., Bera, K., Patil, P. D., Gupta, A., Jain, P., Alilou, M., Khorrami, M., Velcheti, V., Madabhushi, A. Novel, non-invasive imaging approach to identify patients with advanced non-small cell lung cancer at risk of hyperprogressive disease with immune checkpoint blockade. J. Immunother. Cancer 2020, 8, e001343; https://doi.org/10.1136/jitc-2020-001343.Search in Google Scholar PubMed PubMed Central

3. Gristina, V., Malapelle, U., Galvano, A., Pisapia, P., Pepe, F., Rolfo, C., Tortorici, S., Bazan, V., Troncone, G., Russo, A. The significance of epidermal growth factor receptor uncommon mutations in non-small cell lung cancer: a systematic review and critical appraisal. Cancer Treat Rev. 2020, 85, 1; https://doi.org/10.1016/j.ctrv.2020.101994.Search in Google Scholar PubMed

4. Zahavi, D., Weiner, L. Monoclonal antibodies in cancer therapy. Antibodies 2020, 34, 1; https://doi.org/10.3390/antib9030034.Search in Google Scholar PubMed PubMed Central

5. Huang, L., Jiang, S., Shi, Y. Tyrosine kinase inhibitors for solid tumors in the past 20 years (2001–2020). J. Hematol. Oncol. 2020, 13, 1; https://doi.org/10.1186/s13045-020-00977-0.Search in Google Scholar PubMed PubMed Central

6. Liang, Y., Zhang, T., Zhang, J. Natural tyrosine kinase inhibitors acting on the epidermal growth factor receptor: their relevance for cancer therapy. Pharmacol. Res. 2020, 161, 1; https://doi.org/10.1016/j.phrs.2020.105164.Search in Google Scholar PubMed

7. Vallis, K. A., Reilly, R. M., Chen, P., Oza, A., Hendler, A., Cameron, R., Hershkop, M., Iznaga-Escobar, N., Ramos-Suzarte, M., Keane, P. A phase I study of 99mTc-hR3 (DiaCIM), a humanized immunoconjugate directed towards the epidermal growth factor receptor. Nucl. Med. Commun. 2002, 23, 1155; https://doi.org/10.1097/00006231-200212000-00002.Search in Google Scholar PubMed

8. Torres, L. A., Perera, A., Batista, J. F., Hernandez, A., Crombet, T., Ramos, M., Neninger, E., Perez, M., Sanchez, E. L., Romero, S., Aguilar, V., Coca, M. A., Iznaga-Escobar, N. Phase I/II clinical trial of the humanized anti-EGF-r monoclonal antibody h-R3 labeled with 99mTc in patients with tumour of epithelial origin. Nucl. Med. Commun. 2005, 26, 1049; https://doi.org/10.1097/00006231-200512000-00002.Search in Google Scholar PubMed

9. Shih, Y. H., Peng, C. L., Lee, S. Y., Chiang, P. F., Yao, C. J., Lin, W. J., Luo, T. Y., Shieh, M. J. 111In-cetuximab as a diagnostic agent by accessible epidermal growth factor (EGF) receptor targeting in human metastatic colorectal carcinoma. Oncotarget 2015, 18, 16601; https://doi.org/10.18632/oncotarget.3968.Search in Google Scholar PubMed PubMed Central

10. Xiaoxia, W., Qing-Ping, W., Shi, K., Lee, E., Chusilp, C., Abraham, S. D., Sidney, W., Chun, L. Conjugation with 111In-DTPA-poly (ethylene glycol) improves imaging of anti-EGF receptor antibody C225. J. Nucl. Med. 2001, 42, 1530.Search in Google Scholar

11. van Dijk, L. K., Hoeben, B. A., Kaanders, J. H., Franssen, G. M., Boerman, O. C., Bussink, J. Imaging of epidermal growth factor receptor expression in head and neck cancer with SPECT/CT and 111In-labeled cetuximab-F(ab’)2. J. Nucl. Med. 2013, 54, 2118; https://doi.org/10.2967/jnumed.113.123612.Search in Google Scholar PubMed

12. Chen, W., Shen, B., Sun, X. Analysis of progress and challenges of EGFR-targeted molecular imaging in cancer with a focus on affibody molecules. Mol. Imag. 2019, 18, 1; https://doi.org/10.1177/1536012118823473.Search in Google Scholar PubMed PubMed Central

13. Liu, Y., Jin, C., Yang, S., Jia, B., Wang, F., Liu, Z. 125I/111In labeled anti-EGFR monoclonal antibody panitumumab: preparation and biodistribution evaluation in normal mice. J. Nucl. Radiochem. 2010, 32, 362.Search in Google Scholar

14. Boland, W., Bebb, G. The emerging role of nimotuzumab in the treatment of non-small cell lung cancer. Biologics 2010, 9, 289; https://doi.org/10.2147/BTT.S8617.Search in Google Scholar PubMed PubMed Central

15. Ju, Y., Wang, J., Sun, S., Jiao, S. Nimotuzumab treatment and outcome analysis in patients with leptomeningeal metastasis from nonsmall cell lung cancer. J. Cancer Res. Therapeut. 2016, 12, C181; https://doi.org/10.4103/0973-1482.200596.Search in Google Scholar PubMed

16. Yang, Y., Zhou, W., Wu, J., Yao, L., Xue, L., Zhang, Q., Wang, Z., Wang, X., Dong, S., Zhao, J., Yin, D. Antitumor activity of nimotuzumab in combination with cisplatin in lung cancer cell line A549 in vitro. Oncol. Lett. 2018, 15, 5280; https://doi.org/10.3892/ol.2018.7923.Search in Google Scholar PubMed PubMed Central

17. Cai, W. Q., Zeng, L. S., Wang, L. F., Wang, Y. Y., Cheng, J. T., Zhang, Y., Han, Z. W., Zhou, Y., Huang, S. L., Wang, X. W., Peng, X. C., Xiang, Y., Ma, Z., Cui, S. Z., Xin, H. W. The latest battles between EGFR monoclonal antibodies and resistant tumor cells. Front. Oncol. 2020, 24, 1249; https://doi.org/10.3389/fonc.2020.01249.Search in Google Scholar PubMed PubMed Central

18. Yamaguchi, A., Achmad, A., Hanaoka, H., Heryanto, Y. D., Bhattarai, A., Khongorzul, E., Shintawati, R., Kartamihardja, A. A., Kanai, A., Sugo, Y., Ishioka, N. S. Immuno-PET imaging for non-invasive assessment of cetuximab accumulation in non-small cell lung cancer. BMC Cancer 2019, 19, 1; https://doi.org/10.1186/s12885-019-6238-4.Search in Google Scholar PubMed PubMed Central

19. Shah, S. Q., Raana, G. Synthesis and preclinical investigation of 99mTc-p-SCN-Bzl-DTPA-cetuximab for targeting EGFR using head and neck squamous cell carcinoma (HNSCC) xenografts. Mol. Biol. Rep. 2019, 46, 1675; https://doi.org/10.1007/s11033-019-04616-x.Search in Google Scholar PubMed

20. Hoeben, B. A., Molkenboer-Kuenen, J. D., Oyen, W. J., Peeters, W. J., Kaanders, J. H., Bussink, J., Boerman, O. C. Radiolabeled cetuximab: dose optimization for epidermal growth factor receptor imaging in a head-and-neck squamous cell carcinoma model. Int. J. Cancer 2011, 129, 870; https://doi.org/10.1002/ijc.25727.Search in Google Scholar PubMed

21. Shih, Y. H., Peng, C. L., Lee, S. Y., Chiang, P. F., Yao, C. J., Lin, W. J., Luo, T. Y., Shieh, M. J. 111In-cetuximab as a diagnostic agent by accessible epidermal growth factor (EGF) receptor targeting in human metastatic colorectal carcinoma. Oncotarget 2015, 6, 16601; https://doi.org/10.18632/oncotarget.3968.Search in Google Scholar PubMed PubMed Central

22. Hartimath, S. V., El-Sayed, A., Makhlouf, A., Bernhard, W., Gonzalez, C., Hill, W., Parada, A. C., Barreto, K., Geyer, C. R., Fonge, H. Therapeutic potential of nimotuzumab PEGylated-maytansine antibody drug conjugates against EGFR positive xenograft. Oncotarget 2019, 10, 1031; https://doi.org/10.18632/oncotarget.26613.Search in Google Scholar PubMed PubMed Central

23. Fasih, A., Fonge, H., Cai, Z., Leyton, J. V., Tikhomirov, I., Done, S. J., Reilly, R. M. 111In-Bn-DTPA-nimotuzumab with/without modification with nuclear translocation sequence (NLS) peptides: an Auger electron-emitting radioimmunotherapeutic agent for EGFR-positive and trastuzumab (Herceptin)-resistant breast cancer. Breast Cancer Res. Treat. 2012, 135, 189; https://doi.org/10.1007/s10549-012-2137-y.Search in Google Scholar PubMed

24. Goldenberg, A., Masui, H., Divgi, C., Kamrath, H., Pentlow, K., Mendelsohn, J. Imaging of human tumor xenografts with an indium-111-labeled anti-epidermal growth factor receptor monoclonal antibody. J. Natl. Cancer Inst. 1989, 81, 1616; https://doi.org/10.1093/jnci/81.21.1616.Search in Google Scholar PubMed

25. Song, I. H., Noh, Y., Kwon, J., Jung, J. H., Lee, B. C., Kim, K. I., Lee, Y. J., Kang, J. H., Rhee, C. S., Lee, C. H., Lee, T. S., Choi, I. J. Immuno-PET imaging based radioimmunotherapy in head and neck squamous cell carcinoma model. Oncotarget 2017, 8, 92090; https://doi.org/10.18632/oncotarget.20760.Search in Google Scholar PubMed PubMed Central

Received: 2021-05-11

Accepted: 2021-10-05

Published Online: 2021-10-20

Published in Print: 2021-12-20

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Articles in the same Issue

https://doi.org/10.1515/ract-2021-1054

Keywords for this article

111In labeled h-R; biodistribution ad imaging; EGFR; non-small cell lung carcinoma (NSCLC)