A lyophilized peptide radiopharmaceutical kit to target neurotensin receptor for tumor imaging

Document Type: Original Article

Authors

1 Nuclear Science and Technology Research Institute, Atomic Energy Organization of Iran (AEOI), Tehran, Iran

2 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

Abstract

Introduction: Neurotensin (NT) is a tridecapeptide that binds specifically to neurotensin receptors. Several forms of cancer, including small cell lung cancer, colon, pancreatic and prostate carcinomas especially exocrine pancreatic carcinomas express receptors for neurotensin peptide. Radiolabeled neurotensin derivatives with a high affinity for these receptors might be used for scintigraphy. The aim of this work was to prepare a freeze-dried kit formulation for routine preparation of 99mTc labeled neurotensin in nuclear medicine center. Methods: A freeze dried kit containing reducing agent, coligands and HYNIC-Neurotensin derivative for labeling with 99mTc was prepared. Labeling was performed at 95°C for 15 min and radiochemical analysis involved ITLC and HPLC methods. The stability of radiopeptide was checked in the presence of human serum at 37°C up to 24 h. The receptor bound internalization rate was studied in neurotensin receptor expressing HT-29 cells. Biodistribution of radiopeptide was studied in mice. Results: Labeling yield of >95% was obtained corresponding to a specific activity of 80 MBq/nmol. Prepared radioconjugate was stable in human serum and more than 12% of activity was specifically internalized into HT-29 cells up to 4 h. Biodistribution study showed a rapid blood clearance, with renal excretion and specific binding towards NT receptor-positive tissues such as intestines (0.99 ± 0.16% ID/g at 1 h). Conclusion: The favorable characteristics of our new designed labeled peptide formulation make it as a promising candidate for diagnosis of malignant tumors.

Keywords

Main Subjects


Faintuch BL, Teodoro R, Duatti A, Muramoto E, Faintuch S, Smith CJ. Radiolabeled bombesin analogs for prostate cancer diagnosis: preclinical studies. Nucl Med Biol. 2008 May;35(4):401-11.

Reubi JC, Mäcke HR, Krenning EP. Candidates for peptide receptor radiotherapy today and in the future. J Nucl Med. 2005 Jan;46 Suppl 1:67S-75S.

Morinville A, Martin S, Lavallée M, Vincent JP, Beaudet A, Mazella J.  Internalization and trafficking of neurotensin via NTS3 receptors in HT29 cells. Int J Biochem Cell Biol. 2004 Nov;36(11):2153-68.

Kitabgi P. Targeting neurotensin receptors with agonists and antagonists for therapeutic purposes. Curr Opin Drug Discov Devel. 2002 Sep;5(5):764-76.

Vincent JP, Mazella J, Kitabgi P. Neurotensin and neurotensin receptors. Trends Pharmacol Sci. 1999 Jul;20(7):302-9.

Reubi JC, Waser B, Friess H, Büchler M, Laissue J. Neurotensin receptors: a new marker for human ductal pancreatic adenocarcinoma. Gut. 1998 Apr;42(4):546-50.

Oyen WJ, Bodei L, Giammarile F, Maecke HR, Tennvall J, Luster M, Brans B. Targeted therapy in nuclear medicine--current status and future prospects. Ann Oncol. 2007 Nov;18(11):1782-92.

Schubiger PA, Allemann-Tannahill L, Egli A, Schibli R, Alberto R, Carrel-Rémy N, Willmann M, Bläuenstein P, Tourwé D. Catabolism of neurotensins.  Catabolism of neurotensins. Implications for the design of radiolabeling strategies of peptides. Q J Nucl Med. 1999 Jun;43(2):155-8.

Teodoro R, Faintuch BL, Núñez EG, Queiróz RG. Neurotensin(8-13) analogue: radiolabeling and biological evaluation using different chelators. Nucl Med Biol. 2011 Jan;38(1):113-20.

Maina T, Nikolopoulou A, Stathopoulou E, Galanis AS, Cordopatis P, Nock BA. [99mTc]Demotensin 5 and 6 in the NTS1-R-targeted imaging of tumours: synthesis and preclinical results. Eur J Nucl Med Mol Imaging. 2007 Nov;34(11):1804-14.

García-Garayoa E, Maes V, Bläuenstein P, Blanc A, Hohn A, Tourwé D, Schubiger PA. Double-stabilized neurotensin analogues as potential radiopharmaceuticals for NTR-positive tumors. Nucl Med Biol. 2006 May;33(4):495-503.

Abrams MJ, Juweid M, tenKate CI, Schwartz DA, Hauser MM, Gaul FE, Fuccello AJ, Rubin RH, Strauss HW, Fischman AJ. Technetium-99m-human polyclonal IgG radiolabeled via the hydrazino nicotinamide derivative for imaging focal sites of infection in rats. J Nucl Med. 1990 Dec;31(12):2022-8.

Gandomkar M, Najafi R, Shafiei M, Mazidi M, Ebrahimi SE. Preclinical evaluation of [99mTc/EDDA/tricine/HYNIC0, 1-Nal3, Thr8]-octreotide as a new analogue in the detection of somatostatin-receptor-positive tumors. Nucl Med Biol. 2007 Aug;34(6):651-7.

Erfani M, Shafiei M, Mazidi M, Goudarzi M. Preparation and biological evaluation of [(99m)Tc/EDDA/Tricine/HYNIC(0), BzThi(3)]-octreotide for somatostatin receptor-positive tumor imaging. Cancer Biother Radiopharm. 2013 Apr;28(3):240-7.

Sadeghzadeh N, Gandomkar M, Najafi R, Shafiei M, Sadat Ebrahimi SE, Shafiee A, Larijani B. Preparation and evaluation of a new 99mTc labeled bombesin derivative for tumor imaging. J Radioanal Nucl Chem. 2010;283:181-87.

Sadeghzadeh N,  Gandomkar M, Shafiee M, Mazidi M, Goudarzi M, Mirfallah SM, Sadat Ebrahimi SE. Synthesis and evaluation of a new radiolabeled bombesin analogue for diagnosis of GRP receptor expressing tumors. Iran J Nucl Med. 2009;17(1):18-26.

Shirmardi SP, Gandomkar M, Mazidi M, Shafiei M, Ghannadi Maragheh M Synthesis and evaluation of a new bombesin analog labeled with 99mTc as a GRP receptor imaging agent. J Radioanal Nucl Chem. 2011;288:327-35.

Shirmardi SP, Gandomkar M, Maragheh MG, Shamsaei M. Preclinical evaluation of a new bombesin analog for imaging of gastrin-releasing peptide receptors. Cancer Biother Radiopharm. 2011 Jun;26(3):309-16.

Liu S, Edwards DS, Looby RJ, Poirier MJ, Rajopadhye M, Bourque JP, Carroll TR. Labeling cyclic glycoprotein IIb/IIIa receptor antagonists with 99mTc by the preformed chelate approach: effects of chelators on properties of [99mTc]chelator-peptide conjugates. Bioconjug Chem. 1996 Mar-Apr;7(2):196-202.

Kasina S, Rao TN, Srinivasan A, Sanderson JA, Fitzner JN, Reno JM, Beaumier PL, Fritzberg AR. Development and biologic evaluation of a kit for preformed chelate technetium-99m radiolabeling of an antibody Fab fragment using a diamide dimercaptide chelating agent. J Nucl Med. 1991 Jul;32(7):1445-51.

Maina T, Stolz B, Albert R, Bruns C, Koch P, Mäcke H.  Synthesis, radiochemistry and biological evaluation of a new somatostatin analogue (SDZ 219-387) labelled with technetium-99m. Eur J Nucl Med. 1994 May;21(5):437-44.

Liu S, Edwards DS, Looby RJ, Harris AR, Poirier MJ, Barrett JA, Heminway SJ, Carroll TR. Labeling a hydrazino nicotinamide-modified cyclic IIb/IIIa receptor antagonist with 99mTc using aminocarboxylates as coligands. Bioconjug Chem. 1996 Jan-Feb;7(1):63-71. 

Gabriel M, Froehlich F, Decristoforo C, Ensinger C, Donnemiller E, von Guggenberg E, Heute D, Moncayo R. 99mTc-EDDA/HYNIC-TOC and (18)F-FDG in thyroid cancer patients with negative (131)I whole-body scans. Eur J Nucl Med Mol Imaging. 2004 Mar;31(3):330-41.

Babich JW, Fischman AJ. Effect of "co-ligand" on the biodistribution of 99mTc-labeled hydrazino nicotinic acid derivatized chemotactic peptides. Nucl Med Biol. 1995 Jan;22(1):25-30.

Nock BA, Nikolopoulou A, Reubi JC, Maes V, Conrath P, Tourwé D, Maina T.  Toward stable N4-modified neurotensins for NTS1-receptor-targeted tumor imaging with 99mTc. J Med Chem. 2006 Jul 27;49(15):4767-76.

Tanaka K, Masu M, Nakanishi S. Structure and functional expression of the cloned rat neurotensin receptor. Neuron. 1990 Jun;4(6):847-54.

Mulè F, Serio R, Postorino A, Vetri T, Bonvissuto F. Antagonism by SR 48692 of mechanical responses to neurotensin in rat intestine. Br J Pharmacol. 1996 Feb;117(3):488-492.