Re and Tc tricarbonyl complexes: from the suppression of NO biosynthesis in macrophages to in vivo targeting of inducible nitric oxide synthase.
The in vivo molecular imaging of nitric oxide synthase (NOS), the enzyme responsible for the catalytic oxidation of l-arginine to citrulline and nitric oxide (NO), by noninvasive modalities could provide valuable insights into NO/NOS-related diseases. Aiming at the design of innovative (⁹⁹m)Tc(I) complexes for targeting inducible NOS (iNOS) in vivo by SPECT imaging, herein we describe a set of novel (⁹⁹m)Tc(CO)₃ complexes (2-5) and the corresponding rhenium surrogates (2a-5a) containing the NOS inhibitor N(ω)-nitro-l-arginine. The latter is linked through its α-NH₂ or α-COOH group and an alkyl spacer of variable length to the metal center. The complexes 2a (propyl spacer) and 3a (hexyl spacer), in which the α-NH₂ group of the inhibitor is involved in the conjugation to the metal center, presented remarkable affinity for purified iNOS, being similar to that of the free nonconjugated inhibitor (K(i) = 3-8 μM) in the case of 3a (K(i) = 6 μM). 2a and 3a are the first examples of organometallic complexes that permeate through RAW 264.7 macrophage cell membranes, interacting specifically with the target enzyme, as confirmed by the suppression of NO biosynthesis in LPS-treated macrophages (2a, ca. 30% inhibition; 3a, ca. 50% inhibition). The (⁹⁹m)Tc(I)-complexes 2 and 3, stable both in vitro and in vivo, also presented the ability to cross cell membranes, as demonstrated by internalization studies in the same cell model. The biodistribution studies in LPS-pretreated mature female C57BL6 mice have shown that 2 presented an overall higher uptake in most tissues of the LPS-treated mice compared to the control group (30 min postinjection). This increase is significant in lung (3.98 ± 0.63 vs to 0.99 ± 0.13%ID/g), which is known to be the organ with the highest iNOS expression after LPS treatment. These results suggest that the higher uptake in that organ may be related to iNOS upregulation.
Unidade de Ciências Químicas e Radiofarmacêuticas, ITN, Estrada Nacional 10, 2686-953 Sacavém, Portugal.
Bruno L
BL
Oliveira
Paula D
PD
Raposinho
Filipa
F
Mendes
Flávio
F
Figueira
Isabel
I
Santos
António
A
Ferreira
Carlos
C
Cordeiro
Ana P
AP
Freire
João D G
JD
Correia
D000818:Animals; D001120:Arginine / Q000378:metabolism; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D002956:Citrulline / Q000378:metabolism; D056831:Coordination Complexes / Q000737:chemistry / Q000493:pharmacokinetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D015335:Molecular Probes / Q000737:chemistry / Q000493:pharmacokinetics*; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000378:metabolism; D009928:Organ Specificity; D012211:Rhenium / Q000378:metabolism; D013667:Technetium / Q000378:metabolism; D014018:Tissue Distribution
D056831:Coordination Complexes; D004791:Enzyme Inhibitors; D008070:Lipopolysaccharides; D015335:Molecular Probes; D002956:Citrulline; D009569:Nitric Oxide; D012211:Rhenium; D013667:Technetium; D001120:Arginine; D052247:Nitric Oxide Synthase Type II