Preclinical evaluation of somatostatin analogs bearing two macrocyclic chelators for high specific activity labeling with radiometals

Abstract

Radiometallated analogues of the regulatory peptide somatostatin are of interest in the in vivo localization and targeted radiotherapy of somatostatin receptor-overexpressing tumors. An important aspect of their use in vivo is a fast and efficient labeling (complexation) protocol for radiometals along with a high specific activity. We describe in this manuscript synthetic methods for the coupling of two chelators (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid=DOTA) to the bioactive peptide [Tyr3, Thr8]-octreotide (TATE) in order to increase the specific activity (radioactivity in Bq per mole peptide). The full chelator-linker-peptide conjugate was assembled on solid support using standard Fmoc chemistry. Two DOTA-chelators were linked to the peptide using lysine or N,N′-bis(3-aminopropyl)-glycine (Apg); in addition, pentasarcosine (Sar5) was used as a spacer between the chelators and the peptide to probe its influence on biology and pharmacology. Complexation rates with In3+ and Y3+ salts and the corresponding radiometals were high, the bis-DOTA-derivatives showed higher complexation rates and gave higher specific activity than DOTA-TATE. Pharmacological and biological data of the complexed molecules did not show significant differences if compared to the parent peptide [111/natIn-DOTA]-TATE except for [(111/natIn-DOTA)2-Apg]-TATE which showed a lower binding affinity and rate of internalization into tumor cells. The biodistribution of [(111/natIn-DOTA)-Lys(111/natIn-DOTA)]-TATE in the rat tumor model (AR4-2J) showed a high and specific (as shown by a blocking experiment) tracer uptake in somatostatin receptor-positive tissue but a lower tumor uptake compared to [111/natIn-DOTA]-TATE.