Characterization of a novel iodinated sigma-2 receptor ligand as a cell proliferation marker

Overexpression of sigma-2 receptors in human tumors, such as melanoma, breast cancer, small cell lung carcinoma and prostate cancer, has been reported. Furthermore, the expression of sigma-2 receptors parallels the proliferative status of breast tumors implanted in nude mice. Thus, radiolabeled probes with a high affinity and high selectivity targeting sigma-2 receptors may be useful as tumor imaging agents. A conformationally flexible benzamide derivative, 5-bromo-2,3-dimethoxy-N-[2-(6,7-dimethoxy-3,4-dihydro-1H-isoquinolin-2-yl)-butyl]-benzamide, displayed greater than 1000-fold selectivity for sigma-2 receptors (Ki=8.2 and 12,900 nM for sigma-2 and sigma-1, respectively). The corresponding radioiodinated ligand, 5-iodo-2,3-dimethoxy-N-[2-(6,7-dimethoxy-3,4-dihydro-1H-isoquinolin-2-yl)-butyl]-benzamide ([125I]/[123I]I), was successfully prepared via an iododestannylation reaction. Binding studies carried out in membrane homogenates prepared from the mouse mammary tumor cell line (EMT6) with [125I]I showed the desired high binding affinity and high capacity (Kd=0.68±0.06 nM, Bmax=1005±46 fmol/mg protein). The sigma-2-like pharmacological profile of [125I]I binding sites was confirmed by competition studies. Similar binding parameters were also found in EMT6 xenografts (Day 10 and Day 19 implants) from BALB/c mice (Kd=0.43–1.1 nM, Bmax=2025–4528 fmol/mg protein). It was determined by dissection and microSPECT imaging that [125I]/[123]I accumulated in EMT6 tumors established in BALB/c mice (Day 10 implants). Two hours after the tracer injection, the dissection ratio of EMT6 tumor to background (muscle) reached 6–7. However, microSPECT imaging could not clearly delineate the tumors, while the specific localization could be confirmed by ex vivo autoradiography. In summary, the novel iodinated ligand with high affinity and considerable selectivity for sigma-2 receptors may provide a useful tool to characterize sigma-2 receptors in vitro. Further modification of the ligand and the imaging parameters will be needed to improve the signal for in vivo detection.