Peptide-chelating agent conjugate for selective targeting of somatostatin receptor type 1: Synthesis and characterization

Rosalba Mansi, Diego Tesauro, Antonia De Capua, Roberto Fattorusso, Corradina Caracò, Luigi Aloj, Ettore Benedetti, Giancarlo Morelli

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Previously reported results suggest that the analogue of the somatostatin des-AA 1,2,5[D-Trp 8,IAmp 9]-somatostatin (CH-275) peptide bearing chelating agents able to coordinate radioactive metals could be used for scintigraphic imaging of tumor lesions overexpressing sstr1. An efficient synthetic procedure for the preparation of the somatostatin analogue CH-275 and its conjugate DTPAGlu-Gly-CH-275, bearing the chelating agent DTPAGlu (DTPAGlu = N,N-bis[2-[bis(carboxy-ethyl)amino]ethyl]-L-glutamic acid) on the N-terminus, by solid-phase peptide synthesis and 9-flourenymethyoxycarbonyl (Fmoc) chemistry, is here reported. Rapid and efficient labeling of DTPAGlu-Gly-CH-275 was achieved by addition of IIIIn(III) to the compound. Typical yields were greater than 97% as determined by reversed phase high performance liquid chromatography (HPLC) at specific activities in the range 4-9 GBq/μmol (100-250 Ci/mmol). A preliminary biological assay of the binding ability of IIIIn-DTPAGlu-Gly-CH-275 indicates, however, that the labeled compound does not display any specific interaction with somatostatin sstrl receptors in the tested cell lines. To confirm this unexpected negative result, competition binding experiments were carried out, in which fixed tracer amounts of the 125!-labeled somatostatin-14 were incubated with the receptor-expressing cells in the presence of DTPAGlu-Gly-CH-275 or CH-275 at concentrations ranging from 10 -10 to 10 -3 M. While CH-275 shows a IC 50 of 80 nM similar to that already found in displacement experiments on CHO-Kl sstrl-transfected cells, DTPAGlu-Gly-CH-275 displays instead very low or negligible affinity towards this receptor. The NMR solution characterization indicates that the presence of DTPAGlu does not influence the conformational and chemical features of the peptide moiety, thus suggesting that the loss in binding activity should be due to steric hindrance of either the chelating agent DTPAGlu or its indium complex.

Original languageEnglish
Pages (from-to)527-534
Number of pages8
JournalBiopolymers
Volume76
Issue number6
DOIs
Publication statusPublished - 2004

Fingerprint

Bearings (structural)
Chelating Agents
Chelation
Peptides
Somatostatin
High performance liquid chromatography
Indium
Labeling
Tumors
Assays
Experiments
Display devices
Cells
Nuclear magnetic resonance
Imaging techniques
Acids
Metals
somatostatin receptor type 1
CH 275
Somatostatin Receptors

Keywords

  • Indium complexes
  • NMR
  • Nuclear medicine techniques
  • Somatostatin analogue

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics

Cite this

Peptide-chelating agent conjugate for selective targeting of somatostatin receptor type 1 : Synthesis and characterization. / Mansi, Rosalba; Tesauro, Diego; De Capua, Antonia; Fattorusso, Roberto; Caracò, Corradina; Aloj, Luigi; Benedetti, Ettore; Morelli, Giancarlo.

In: Biopolymers, Vol. 76, No. 6, 2004, p. 527-534.

Research output: Contribution to journalArticle

Mansi, Rosalba ; Tesauro, Diego ; De Capua, Antonia ; Fattorusso, Roberto ; Caracò, Corradina ; Aloj, Luigi ; Benedetti, Ettore ; Morelli, Giancarlo. / Peptide-chelating agent conjugate for selective targeting of somatostatin receptor type 1 : Synthesis and characterization. In: Biopolymers. 2004 ; Vol. 76, No. 6. pp. 527-534.
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AU - Mansi, Rosalba

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AU - De Capua, Antonia

AU - Fattorusso, Roberto

AU - Caracò, Corradina

AU - Aloj, Luigi

AU - Benedetti, Ettore

AU - Morelli, Giancarlo

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N2 - Previously reported results suggest that the analogue of the somatostatin des-AA 1,2,5[D-Trp 8,IAmp 9]-somatostatin (CH-275) peptide bearing chelating agents able to coordinate radioactive metals could be used for scintigraphic imaging of tumor lesions overexpressing sstr1. An efficient synthetic procedure for the preparation of the somatostatin analogue CH-275 and its conjugate DTPAGlu-Gly-CH-275, bearing the chelating agent DTPAGlu (DTPAGlu = N,N-bis[2-[bis(carboxy-ethyl)amino]ethyl]-L-glutamic acid) on the N-terminus, by solid-phase peptide synthesis and 9-flourenymethyoxycarbonyl (Fmoc) chemistry, is here reported. Rapid and efficient labeling of DTPAGlu-Gly-CH-275 was achieved by addition of IIIIn(III) to the compound. Typical yields were greater than 97% as determined by reversed phase high performance liquid chromatography (HPLC) at specific activities in the range 4-9 GBq/μmol (100-250 Ci/mmol). A preliminary biological assay of the binding ability of IIIIn-DTPAGlu-Gly-CH-275 indicates, however, that the labeled compound does not display any specific interaction with somatostatin sstrl receptors in the tested cell lines. To confirm this unexpected negative result, competition binding experiments were carried out, in which fixed tracer amounts of the 125!-labeled somatostatin-14 were incubated with the receptor-expressing cells in the presence of DTPAGlu-Gly-CH-275 or CH-275 at concentrations ranging from 10 -10 to 10 -3 M. While CH-275 shows a IC 50 of 80 nM similar to that already found in displacement experiments on CHO-Kl sstrl-transfected cells, DTPAGlu-Gly-CH-275 displays instead very low or negligible affinity towards this receptor. The NMR solution characterization indicates that the presence of DTPAGlu does not influence the conformational and chemical features of the peptide moiety, thus suggesting that the loss in binding activity should be due to steric hindrance of either the chelating agent DTPAGlu or its indium complex.

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