We have developed
a diphosphine (DP) platform for radiolabeling
peptides with 99mTc and 64Cu for molecular SPECT
and PET imaging, respectively. Two diphosphines, 2,3-bis(diphenylphosphino)maleic
anhydride (DPPh) and 2,3-bis(di-p-tolylphosphino)maleic
anhydride (DPTol), were each reacted with a Prostate Specific
Membrane Antigen-targeted dipeptide (PSMAt) to yield the bioconjugates
DPPh-PSMAt and DPTol-PSMAt, as well as an integrin-targeted
cyclic peptide, RGD, to yield the bioconjugates DPPh-RGD
and DPTol-RGD. Each of these DP-PSMAt conjugates formed
geometric cis/trans-[MO2(DPX-PSMAt)2]+ (M = 99mTc, 99gTc, natRe; X = Ph, Tol) complexes when
reacted with [MO2]+ motifs. Furthermore, both
DPPh-PSMAt and DPTol-PSMAt could be formulated
into kits containing reducing agent and buffer components, enabling
preparation of the new radiotracers cis/trans-[99mTcO2(DPPh-PSMAt)2]+ and cis/trans-[99mTcO2(DPTol-PSMAt)2]+ from aqueous 99mTcO4
– in 81% and 88% radiochemical yield (RCY), respectively, in 5 min
at 100 °C. The consistently higher RCYs observed for cis/trans-[99mTcO2(DPTol-PSMAt)2]+ are attributed
to the increased reactivity of DPTol-PSMAt over DPPh-PSMAt. Both cis/trans-[99mTcO2(DPPh-PSMAt)2]+ and cis/trans-[99mTcO2(DPTol-PSMAt)2]+ exhibited
high metabolic stability, and in vivo SPECT imaging
in healthy mice revealed that both new radiotracers cleared rapidly
from circulation, via a renal pathway. These new diphosphine bioconjugates
also furnished [64Cu(DPX-PSMAt)2]+ (X = Ph, Tol) complexes rapidly, in a high RCY (>95%),
under
mild conditions. In summary, the new DP platform is versatile: it
enables straightforward functionalization of targeting peptides with
a diphosphine chelator, and the resulting bioconjugates can be simply
radiolabeled with both the SPECT and PET radionuclides, 99mTc and 64Cu, in high RCYs. Furthermore, the DP platform
is amenable to derivatization to either increase the chelator reactivity
with metallic radioisotopes or, alternatively, modify the radiotracer
hydrophilicity. Functionalized diphosphine chelators thus have the
potential to provide access to new molecular radiotracers for receptor-targeted
imaging.