Radiolabeling morpholinos with (90)Y, (111)In, (188)Re and (99m)Tc

This laboratory is investigating morpholinos (MORF), a DNA analogue, for radiopharmaceutical applications. While we routinely radiolabel with (99m)Tc, we have now labeled MORFs with (111)In, (188)Re and (90)Y in anticipation of therapeutic studies. METHODS: A 25 mer MORF with a primary amine on the 3' equivalent end attached via a 10 member linker was conjugated with an isothiocyanate backbone derivative of DOTA (for labeling with (111)In and (90)Y) and with NHS-MAG(3) (for labeling with (188)Re and (99m)Tc). The in vitro stability of labeled MORFs were investigated and biodistribution was carried out in normal mice. RESULTS: As evident by size exclusion HPLC, ITLC and Sep-Pak analysis, all four radiolabeled MORFs were successfully radiolabeled. In each case, the labeled MORFs showed one sharp peak in HPLC that shifted completely to earlier retention times following addition of a polymer conjugated with the complementary MORF. In saline at room temperature and in 37 degrees C serum, the radioactivity profile of (111)In, (188)Re and (99m)Tc was unchanged over 48 h while over the same period, the (90)Y profile showed a pronounced lower molecular weight peak which did not shift and was shown to be most probably due to (90)Y-DOTA resulting from radiolysis. In addition, the recovery of (188)Re on HPLC decreased as samples aged probably due to oxidation to perrhenate which was retained by the HPLC column. The biodistributions at 1, 3 and 6 h in normal mice showed no important differences among all four labels with the exception that levels of radioactivity in stomach and thyroid were higher in the case of (188)Re due to in vivo oxidation of the radiolabel to perrhenate. CONCLUSIONS: When radiolabeled with DOTA, (90)Y-labeled MORF showed increased instabilities relative to that of (111)In and when radiolabeled with MAG(3), (188)Re showed in vitro and in vivo instabilities compared to (99m)Tc, but all labels were still largely intact after 48 h in saline or serum. Possibly because of the rapid clearance of MORFs, no important differences in biodistribution among (90)Y, (111)In and (99m)Tc labels were evident in normal mice. These strategies for labeling MORF with (90)Y and (188)Re therefore appear to be suitable for therapeutic applications although both show some evidence of instabilities.