Effect of using different co-ligands during 99mTc-labeling of J18 peptide on SK-MES-1 cell binding and tumor targeting

Document Type : Original Article


1 Department of Radiopharmacy, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran

2 Cardiovascular Diseases Research Center, Department of Cardiology, Heshmat Hospital, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran

3 Department of Nuclear Medicine and Molecular Imaging, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

4 Department of Nuclear Medicine and Molecular Imaging, Clinical Development Research Unit of Farshchian Heart Center, Hamadan University of Medical Sciences, Hamadan, Iran


Objective(s): Lung cancer is the main cause of cancer death, and its incidence is increasing worldwide. The goal of this study is to evaluate in vitro and in vivo tumor targeting behavior of [99mTc]Tc -HYNIC-(Ser)3-J18 in lung carcinoma (SK-MES-1)-bearing mice.
Materials and Methods: The J18 (RSLWSDFYASASRGP) peptide was conjugated with hydrazinonicotinamide (HYNIC) via three serine amino acids as a linker at the peptide’s N-terminal and then labeled with technetium-99m using tricine and tricine/EDDA as the co-ligands. The radiolabeled peptides were assessed for in vitro receptor binding, specific binding, and saturation affinity. In vivo biodistribution studies were also performed for 99mTc-peptide 1 (tricine co-ligand) and 99mTc-peptide 2 (tricine/EDDA coligands) in nude mice bearing SK-MES-1 xenograft tumors.
Results: In vitro studies showed high specific binding for 99mTc-peptide 1 in SKMES-1 cells compared with 99mTc-peptide 2 (11.5 vs. 4.5). The Kd values for 99mTc-peptide 1 and 99mTc-peptide 2 were reported to be 3.1±0.3 nM and 3.46 ± 0.8 nM, respectively.  The biodistribution study also showed high significant tumor to muscle ratios were 5.1 and 6.18 for 99mTc-peptide 1 at 1 and 2 hr after injection, respectively, while these ratios were 3.81 and 5.18 for peptide 2, respectively.
Conclusion: Overall, 99mTc-labeled J18 peptide in the presence of tricine as co-ligand has better in vitro and in vivo tumor targeting properties in SK-MES-1 cells than tricine/EDDA co-ligands. These findings show that the 99mTc-labeled J18 peptide is a good candidate for lung carcinoma targeting.


1. DaSilva J, Dalstein V, Polette M, Nawrocki-Raby B.  Phenotypical plasticity and targeted therapies in non-small cell lung carcinomas. Rev Mal Respir 2019; 36:438- 441.
2. Herbst RS, Morgensztern D, Boshoff  C. The biology and management of non-small cell lung cancer. Nature 2018; 553:446-454.
3. Taylor SA , Mallett S , Ball S , Beare S, Bhatnagar G, Bhowmik A et al. Diagnostic accuracy of whole-body MRI versus standard imaging pathways for metastatic disease in newly diagnosed non-small-cell lung cancer: the prospective Streamline L trial. Lancet Respir Med 2019; 7: 523–532.
4. Sarinas PS, Chitkara RK . PET and SPECT in the management of lung cancer. Curr Opin Pulm Med 2002; 8: 257-264.
5.  Soendergaard M , Newton-Northup JR , Deutscher SL . In vivo phage display selection of an ovarian cancer targeting peptide for SPECT/CT imaging. Am J Nucl Med Mol Imaging 2014; 4: 561-570.    
6. Shaghaghi Z, Abedi SM, Hosseinimehr SJ. 99mTc‐HYNIC‐(Ser) 3‐J18 peptide: A radiotracer for non‐small‐cell lung cancer targeting. Chem biol drug des 2018; 92:1214-1220.
7.  Shaghaghi Z , Abedi SM , Hosseinimehr SJ. Tricine co-ligand improved the efficacy of 99mTc-HYNIC-(Ser)3- J18 peptide for targeting and imaging of non-small-cell lung cancer. Biomed Pharmacother 2018; 104:325-331.
8.  Liu B , Wu X, Liu B, Wang C, Liu Y, Zhou Q, et al. MiR-26a enhances metastasis potential of lung cancer cells via AKT pathway by targeting PTEN. Biochim Biophys Acta 2012; 1822:1692-1704.
9. Yongchun Z , Linwei T , Xicai W , Lianhua Y , Guangqiang Z , Ming Y, et al  .MicroRNA-195 inhibits non-small cell lung cancer cell proliferation, migration and invasion by targeting MYB. Cancer Lett 2014; 347:65-74.
10. Zahmatkesh MH, Abedi SM, Hosseinimehr SJ.99mTc-HYNIC-D4 Peptide: A New Small Radiolabeled Peptide for Non Small Cell Lung Tumor Targeting. Anticancer Agents Med Chem 2017; 17:734-740.
11. Hadei SK, Alvandi M, Ramezani M, Aloosh O, Shaghaghi Z, Moradi A. Applying Wells score to inconclusive perfusion only modified PIOPED II (Prospective Investigation of Pulmonary Embolism Diagnosis II)  readings in order to optimize the lung scintigraphy diagnostic yield in acute pulmonary Embolism detection. Ann Nucl Med 2020 ; 34 :521-526.
12. Shahsavari S, Shaghaghi Z, Abedi SM, Hosseinimehr SJ. Evaluation of 99mTc-HYNIC-(ser)3-LTVPWY peptide for glioblastoma imaging. Int J Radiat Biol  2020; 96:502-509.
13. Alvandi M, Shaghaghi Z, Aryafar V, Fariba F, Sanaei Z. The evaluation of left ventricular dyssynchrony in hypertensive patients with a preserved systolic function undergoing gated SPECT myocardial perfusion  imaging. Ann Nucl Med 2019;33:899-906.
14.  Farzipour S, Hosseinimehr SJ. Correlation between in vitro and in vivo Data of Radiolabeled Peptide for Tumor Targeting. Mini Rev Med Chem 2019;19 :950-960.
15. Torabizadeh SA, Abedi SM, Noaparast Z, Hosseinimehr SJ. Comparative assessment of a 99mTc labeled   H1299.2-HYNIC peptide bearing two different co-ligands for tumor-targeted imaging. Bioorg Med Chem 2017;25:2583-2592.
16. Okarvi SM. Peptide-based radiopharmaceuticals: future tools for diagnostic imaging of cancers and other diseases. Med Res Rev 2004;24:357–397.
17. Heppeler A, Froidevaux S, Eberle AN, Maecke HR. Receptor targeting for tumor localisation and therapy with radiopeptides. Curr Med Chem 2000;7:971–994.
18. Guo H, Miao Y. Introduction of an 8-aminooctanoic acid linker enhances uptake of 99mTc-labeled lactam bridge-cyclized alpha-MSH peptide in melanoma. J Nucl Med 2014; 55: 2057–2063.
19. de Visser M, Bernard HF, Erion JL et al. Novel 111In-labelled bombesin analogues for molecular imaging of prostate tumours. Eur J Nucl Med Mol Imaging 2007;34: 1228-1238.
20. Lane SR, Nanda P, Rold TL et al. Optimization, biological evaluation and microPET imaging of copper-64-labeled bombesin agonists, (64Cu-NO2A-(X)- BBN(7–14)NH2), in a prostate tumor xenografted mouse model. Nucl Med Biol 2010; 37:751–761.
21. Sun X, Li Y, Liu T, Li Z, Zhang X, Chen X. Peptide-based imaging agents for cancer detection. Adv Drug Deliv Rev. 2017;110-111:38-51.
22. Tyldesley B, Grieve J. Muscles, nerves and movement: in human occupation. 2009. John Wiley & Sons
23.Weinstein-Oppenheimer CR, Blalock WL, Steelman LS, Chang F, McCubrey JA. The Raf signal transduction cascade as a target for chemotherapeutic intervention in growth factor-responsive tumors. Pharmacol Ther 2000; 88:229-279.
24.Ceccarelli S, Romano F, Angeloni A, Marchese C .Potential dual role of KGF/KGFR as a target option in novel   therapeutic strategies for the treatment of cancers and mucosal damages. Expert Opin Ther Targets 2012; 16 :377-393.
25. Torabizadeh SA, Hosseinimehr SJ. The influence of co-ligands on improving tumor targeting of 99mTc-HYNIC conjugated peptides. Mini Rev Med Chem 2017;17:86-94.