Side-Chain Modified [Tc]Tc-DT1 Mimics: A Comparative Study in NTSR-Positive Models.

Radiolabeled neurotensin analogs have been developed as candidates for theranostic use against neurotensin subtype 1 receptor (NTSR)-expressing cancer. However, their fast degradation by two major peptidases, neprilysin (NEP) and angiotensin-converting enzyme (ACE), has hitherto limited clinical success. We have recently shown that palmitoylation at the -amine of Lys in [Tc]Tc-[Lys]DT1 (DT1, N-Gly-Arg-Arg-Pro-Tyr-Ile-Leu-OH, N = 6-(carboxy)-1,4,8,11-tetraazaundecane) led to the fully stabilized [Tc]Tc-DT9 analog, displaying high uptake in human pancreatic cancer AsPC-1 xenografts but unfavorable pharmacokinetics in mice. Aiming to improve the in vivo stability of [Tc]Tc-DT1 without compromising pharmacokinetics, we now introduce three new [Tc]Tc-DT1 mimics, carrying different pendant groups at the -amine of Lys: MPBA (4-(4-methylphenyl)butyric acid)-[Tc]Tc-DT10; MPBA via a PEG4-linker-[Tc]Tc-DT11; or a hydrophilic PEG6 chain-[Tc]Tc-DT12. The impact of these modifications on receptor affinity and internalization was studied in NTSR-positive cells. The effects on stability and AsPC-1 tumor uptake were assessed in mice without or during NEP/ACE inhibition. Unlike [Tc]Tc-DT10, the longer-chain modified [Tc]Tc-DT11 and [Tc]Tc-DT12 were significantly stabilized in vivo, resulting in markedly improved tumor uptake compared to [Tc]Tc-DT1. [Tc]Tc-DT11 was found to achieve the highest AsPC-1 tumor values and good pharmacokinetics, either without or during NEP inhibition, qualifying for further validation in patients with NTSR-positive tumors using SPECT/CT.