title: Peptide-chelating agent conjugate for selective targeting of somatostatin receptor type 1: synthesis and characterization.
abstract: Previously reported results suggest that the analogue of the somatostatin des-AA1,2,5[D-Trp8,IAmp9]-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-fluorenylmethoxycarbonyl (Fmoc) chemistry, is here reported. Rapid and efficient labeling of DTPAGlu-Gly-CH-275 was achieved by addition of 111In(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/micromol (100-250 Ci/mmol). A preliminary biological assay of the binding ability of 111In-DTPAGlu-Gly-CH-275 indicates, however, that the labeled compound does not display any specific interaction with somatostatin sstr1 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 125I-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 IC50 of 80 nM similar to that already found in displacement experiments on CHO-K1 sstr1-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.
authors.affiliation: Department of Biological Chemistry & Interuniversity Research Center on Bioactive Peptides (CIRPeB), University of Naples "Federico II," Via Mezzocannone, 6/8 I-80134 Naples, Italy.
authors.forename: Rosalba
authors.initials: R
authors.lastname: Mansi
authors.forename: Diego
authors.initials: D
authors.lastname: Tesauro
authors.forename: Antonia
authors.initials: A
authors.lastname: De Capua
authors.forename: Roberto
authors.initials: R
authors.lastname: Fattorusso
authors.forename: Corradina
authors.initials: C
authors.lastname: Caracò
authors.forename: Luigi
authors.initials: L
authors.lastname: Aloj
authors.forename: Ettore
authors.initials: E
authors.lastname: Benedetti
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Morelli
mesh_terms: D000595:Amino Acid Sequence; D002614:Chelating Agents / Q000138:chemical synthesis* / Q000737:chemistry*; D007205:Indium Radioisotopes; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry*; D019275:Radiopharmaceuticals / Q000138:chemical synthesis / Q000737:chemistry; D017481:Receptors, Somatostatin / Q000737:chemistry*; D013004:Somatostatin / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry*
chemical_list: C110576:CH 275; D002614:Chelating Agents; D007205:Indium Radioisotopes; D010455:Peptides; D019275:Radiopharmaceuticals; D017481:Receptors, Somatostatin; C101130:somatostatin receptor type 1; D013004:Somatostatin