title: Evaluation of 99mTc-MAG3-annexin V: influence of the chelate on in vitro and in vivo properties in mice.
abstract: We conjugated mercaptoacetyltriglycine (MAG(3)) to rh-annexin V to permit radiolabeling with (99m)Tc in an effort to decrease the high kidney and liver accumulation observed for (99m)Tc-labeled Hynic-annexin V. The 36-kDa protein was conjugated at a 5:1 molar ratio with NHS-MAG(3) in HEPES buffer pH 7.8 at room temperature, then quenched with glycine and purified by dialysis. The biopotency of the resulting MAG(3)-annexin was similar to that of Hynic-annexin as determined by a sensitive red blood cell membrane affinity binding assay and a surface plasmon resonance (SPR) assay. The (99m)Tc radiolabeling of MAG(3)-annexin resulted in radiochemical yields of 90% under mildly basic pH conditions. Biodistribution data in normal mice clearly showed a significant decrease in kidney and liver uptake at 1 h postinjection for the (99m)Tc MAG(3)-annexin compared to the (99m)Tc Hynic-annexin (from 24% ID to 4% ID for the liver, and 45% ID to 15% ID for the kidneys, respectively). Autoradiography of the kidneys showed retention of radioactivity in the collecting tubules following administration of both labeled annexins. The (99m)Tc MAG(3)-annexin biodistribution was also characterized by a lower retention of radioactivity in the whole body, but with small intestine accumulation over fivefold higher than observed with (99m)Tc Hynic-annexin. These findings show a definite improvement in renal and hepatic clearance of the MAG(3) radioligand. However, due to the increased radioactivity uptake in the small intestines, the early in vivo detection of ongoing apoptosis in the lower abdomen might be more difficult with (99m)Tc MAG(3)-annexin. Nevertheless, (99m)Tc MAG(3)-annexin may be an attractive alternative to (99m)Tc Hynic-annexin for the in vivo imaging of phosphatidylserine receptors.
authors.affiliation: Department of Radiology, University of Massachusetts Medical School, Worcester, 01655, USA. jlvand@earthlink.net
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Vanderheyden
authors.forename: Guozheng
authors.initials: G
authors.lastname: Liu
authors.forename: Jiang
authors.initials: J
authors.lastname: He
authors.forename: Bhavesh
authors.initials: B
authors.lastname: Patel
authors.forename: Jonathan F
authors.initials: JF
authors.lastname: Tait
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Hnatowich
mesh_terms: D000818:Animals; D002614:Chelating Agents / Q000737:chemistry; D004353:Drug Evaluation, Preclinical; D008297:Male; D008657:Metabolic Clearance Rate; D051379:Mice; D009842:Oligopeptides / Q000493:pharmacokinetics*; D009928:Organ Specificity; D009942:Organometallic Compounds / Q000493:pharmacokinetics*; D019275:Radiopharmaceuticals / Q000737:chemistry / Q000493:pharmacokinetics; D017263:Technetium Tc 99m Mertiatide / Q000737:chemistry / Q000493:pharmacokinetics*; D014018:Tissue Distribution
chemical_list: D002614:Chelating Agents; D009842:Oligopeptides; D009942:Organometallic Compounds; D019275:Radiopharmaceuticals; C510970:technetium 99m mercaptoacetyltriglycine-annexin V; D017263:Technetium Tc 99m Mertiatide