For targeted radionuclide therapy, radioligands that exhibit high and persistent tumor uptake are indispensable. We previously synthesized a 99mTc-labeled hexavalent RGD peptide (99mTc-(RGD)6) as a tumor imaging agent targeting integrin αvβ3. 99mTc-(RGD)6 showed high in vivo tumor uptake with long retention due to simultaneous binding to multiple integrin αvβ3 receptors. The purpose of this study was to apply this finding to the design of a multivalent RGD peptide labeled with 211At, a promising α-emitting radionuclide for radionuclide therapy. As a candidate compound, a tetravalent RGD peptide (H2N-(RGD)4) was synthesized and radiolabeled with 125I, a homologous element of At, for basic studies. As expected, 125I-(RGD)4 retained the capability of simultaneous binding and showed comparable in vivo tumor uptake to 99mTc-(RGD)6. Finally, 211At-(RGD)4 was synthesized with >95% radiochemical purity and exhibited an almost identical biodistribution pattern to 125I-(RGD)4. These results indicate that 211At-(RGD)4 might be a potential radioligand for integrin αvβ3-targeted radionuclide therapy.

As the "molecule of the century", 2-deoxy-2-[18F]fluoro-d-glucose ([18F]FDG) is a radioactive 18F-labeled glucose derivative with a wide range of applications for positron emission tomography (PET) imaging. Single photon emission computed tomography (SPECT) imaging is widely used, but there is no clinical probe comparable to [18F]FDG. In our previous work, [99mTc]Tc-CN5DG and [99mTc]Tc-CN7DG were successfully developed and achieved high-quality SPECT images. However, they still have the disadvantage of low tumor uptake and/or high uptake by nontarget organs. To develop novel tumor imaging agents with high tumor uptake and excellent tumor/nontarget ratios, in this study, starting from d-glucosamine hydrochloride, four phenyl group-containing isonitrile ligands were designed, synthesized, and radiolabeled with 99mTc. All the complexes had high radiochemical purity and good hydrophilicity and stability. Biodistribution experiments showed that [99mTc]Tc-L4 (i.e., [99mTc]Tc-CNMBDG) had the highest tumor uptake and tumor/background ratios among the four probes. In SPECT imaging studies, the tumor detected by [99mTc]Tc-L4 was more clearly visible than that of [99mTc]Tc-CN7DG because of the inappreciable interference from abdominal uptake. Preliminary clinical studies of [99mTc]Tc-L4 have been conducted and successfully showed the lesion location in a patient with non-small-cell lung cancer. In summary, [99mTc]Tc-L4 is expected to be a promising tumor SPECT imaging agent.

The attachment of an ethyne substituent in the para position of phenylisocyanide, CNPh^pC≡CH, enables the isocyanide to replace carbonyl ligands in the coordination sphere of common technetium(I) starting materials such as (NBu₄)[Tc₂(μ-Cl)₃(CO)₆]. The ligand exchange proceeds under thermal conditions and finally forms the corresponding hexakis(isocyanide)technetium(I) complex. The product undergoes a copper-catalyzed cycloaddition ("Click" reaction), e.g., with benzyl azide, which gives the [Tc(CNPh^azole)₆]⁺ cation. The free, uncoordinated "Click" product is obtained from a reaction of the corresponding tetrakis(CNPh^azole)copper(I) complex and NaCN. It readily reacts with mer-[Tc(CO)₃(tht)(PPh₃)₂](BF₄) (tht = tetrahydrothiophene) under exchange of the thioether ligand. Alternatively, [Cu(CNPh^azole)₄]⁺ can be used as a transmetalation reagent for the synthesis of the hexakis(isocyanide)technetium(I) complex, which is the preferable approach for the synthesis of the technetium complex with the short-lived nuclear isomer ^99mTc, and a corresponding protocol for [^99mTc(CNPh^azole)₆]⁺ is reported. The ⁹⁹Tc and copper complexes have been studied by single-crystal X-ray diffraction and/or spectroscopic methods including IR and multinuclear NMR spectroscopy.

Reactions of the alkyl isocyanide fac-[Tc(CO)₃(CNR)₂Cl] complexes (2) (CNR = CNⁿBu or CN^tBu) with the sterically encumbered isocyanide CNp-FAr^DarF2 [DArF = 3,5-(CF₃)₂C₆H₃] allow a selective exchange of the carbonyl ligands of 2 and the isolation of the mixed-isocyanide complexes mer,trans-[Tc(CNp-FAr^DarF2)₃(CNR)₂Cl] (3). Depending on the steric requirements of the residues R, the remaining chlorido ligand can be replaced by another isocyanide ligand. Cationic complexes such as mer-[Tc(CNp-FAr^DarF2)₃(CNⁿBu)₃]⁺ (4a) or mer,trans-[Tc(CNp-FAr^DarF2)₃(CNⁿBu)₂(CN^tBu)]⁺ (6) have been prepared in this way and isolated as their PF₆^- salts. mer,trans-[Tc(CNp-FAr^DarF2)₃(CNⁿBu)₂(CN^tBu)](PF₆) represents to the best of our knowledge the first transition-metal complex with three different isocyanides in its coordination sphere. Since the degree of the ligand exchange seems to be controlled both by the electronic and steric measures of the incoming isocyanides, we undertook similar reactions with the sterically less demanding p-fluorophenyl isocyanide, CNPh^pF, which indeed readily led to the hexakis(isocyanide)technetium(I) cation through an exchange of all ligands in the staring materials [Tc₂(CO)₆(μ-Cl)₃]^- or fac-[Tc(CO)₃(CNR)₂Cl]. The influence of the substituents at the isocyanide ligands in such reactions has been reasoned with the density functional theory-derived electrostatic potential at the accessible surface of the corresponding isocyanide carbon atoms.

Multivalent RGD peptides have been used as an excellent targeting vector to integrin α_vβ₃-positive tumors. However, little attention has been paid to the influence of linker molecules in multivalent RGD peptides on their dissociation kinetics from tumor cells. In this study, we evaluated the dissociation kinetics of ^99mTc-labeled hexavalent RGD peptides which have (CH₂-CH₂-O)_n (n = 4, [^99mTc][Tc(L1)₆]⁺ and n = 12, [^99mTc][Tc(L2)₆]⁺) or (_DPro-Gly)_n (n = 1, [^99mTc][Tc(L3)₆]⁺; n = 6, [^99mTc][Tc(L4)₆]⁺; and n = 9, [^99mTc][Tc(L5)₆]⁺) as a linker molecule. The results showed that [^99mTc][Tc(L4)₆]⁺ and [^99mTc][Tc(L5)₆]⁺ displayed slower dissociation kinetics and [^99mTc][Tc(L4)₆]⁺ showed exceptionally high in vitro cellular uptake (203.1 ± 16.7% dose/mg protein) and the highest tumor to blood ratio (138.1 ± 26.3 at 4 h p.i.) in tumor bearing nude mice. These findings indicate that the use of appropriate length of (_DPro-Gly)_n would maximize the binding of multivalent RGD peptides to clustered integrin α_vβ₃.